Spider mite silk proteins

US008501913B2
(12) Ulllted States Patent
(10) Patent N0.:
Grbic et al.
(54)
US 8,501,913 B2
(45) Date of Patent:
SPIDER MITE SILK PROTEINS
(56)
Aug. 6, 2013
References Cited
(75) Inventors: Miodrag Grbic, Ontario (CA);
FOREIGN PATENT DOCUMENTS
Vojislava Grbic, Ontario (CA);
W0
W0 91/ 16351
10/1991
Van
Stephane
De Peer’
Rombauts,
Ghent (BE)
Ghent (BE); Yves
W0
WO 2011/039345é
4/2011
(73) Assignees: VIB VZW, Ghent (BE); Universiteit
OTHER PUBLICATIONS
Gent, Ghent (BE); The University of
Western Ontario, London, Ontario (CA)
Craig et al., A comparison of the composition of slk proteins pro
duced by spiders and insects; International Journal of Biological
Subject to any disclaimer, the term of this
GIMP et a1~~ Mity mofieli Tf-flmnychus WHY-"a6, a Candidate for
patent is extended Or adjusted under 35
U S C 154 b b 0 da
chelicerate model organism;B1oEssays29z489-496; 2007. _
Coll1n et al.; Characterrzatron of s1lk spun by the embropteran,
Macromolecules 24 (1999) 109-118.
(*) Notice:
' ' '
( ) y
ys'
Antipaluria urichi; Insect Biochemistry and Molecular Biology; 39
(21)
Appl. N0.:
13/499,890
(22)
PCT Flled:
Oct‘ 1’ 2010
(2009) 75-82.
Collin et al.; Characterization of silk spun by the embiopteran,
Antipaluria urichi; XP002619960; Dec. 16, 2008.
Craig et al., A comparison of the composition of silk proteins pro
duced by spiders and insects; International Journal of Biological
(86)
PCT NO_;
PCT/EP2010/064632
Macromolecules 24 (1999) 109-118.
Grbic et al..; Mity model: Tetranychus urticae, a candidate for
chelicerate model organism; BioEssays29:489-496; 2007.
HaZan et al.; Spider Mite WebbingiIII. SolubiliZation and Amino
§ 371 (C)(1),
(2), (4) Date:
(87)
Jun. 7, 2012
Acid Composition of the Silk Protein; Comp. Biochem. Physiol.,
1975, vol. 51B, pp. 457-462.
International Search Report; PCT/EP2010/064632 dated Feb. 23,
PCT Pub. N0.: WO2011/039345
2011‘
.
Teule et al.; A protocol for the production of recombinant spider
PCT Pub' Date' Apr‘ 7’ 2011
(65)
silklike proteins for arti?cial ?ber spinning; Nature Protocols; vol. 4,
No. 3; 2009; pp. 341-355.
Tian et al.; Analysis of Major Ampullate Silk cDNAs from Two
Pl‘iOl‘ PllbliC?tiOIl Data
NonOrb-Weaving Spiders; Bio Macromolecules; vol. 5, No. 3; May/
Tian et al.; Analysis of Maj or Amullate Silk cDNAs from Two
(30)
Foreign Application Priority Data
Oct. 2, 2009
NonOrb-Weaving Spiders; XP-002619958; Jul. 5, 2004.
(EP) ................................... .. 09172104
Primary Examiner * Manjunath R210
Assistant Examiner * Gerard Lacourciere
(51)
Int Cl
(74) Attorney, Agent, or Firm * TraskBr‘itt
A61K 38/1 7
(2006.01)
C12P 21/06
(2006.01)
(
57
C121) 21/04
(200601)
Described are srlk prote1ns derived from'sprder mite, more
(52) U 5 Cl
)
58
_
_
_
_
_
_
speci?cally derived from Tetranychus urtzcae. More speci?
U'séc '
(
ABSTRACT
)
530/353_ 435/69 1_ 435/71 1
"""""""""""" "
’
' ’
Field of Classi?cation Search
None
See application ?le for complete search history.
'
cally, described is the use ofthese proteins to make ?bers, or
?ber-composed material and the resulting ?bers and materi
a15_
15 Claims, N0 Drawings
US 8,501,913 B2
1
2
SPIDER MITE SILK PROTEINS
silk ?bers for different purposes, such as dragline silk or
major ampullate silk, capture-spiral silk, tubuliform silk,
aciniform silk and minor-ampullate silk.
The most investigated type of spider silk is the dragline or
CROSS-REFERENCE To RELATED
APPLICATION(S)
major ampullate (MA) silk that is secreted by the major
ampullate glands of the spider. The dragline is used to support
This is a national phase entry under 35 USC §371 of
the spider When constructing a Web and to prevent it from
falling. This function results in mechanical properties com
bining a hi ghYoung’ s modulus With a high strength. Due to its
International Patent Application PCT/EP2010/064632, ?led
Oct. 1, 2010, published in English as International Patent
Publication WO 2011/039345 Al on Apr. 7, 2011, Which
claims bene?t under Article 8 of the Patent Cooperation
siZe and accessibility, the major ampullate gland has been the
focus of most studies.
Treaty to European Patent Application Serial No.
091721043, ?led Oct. 2, 2009.
spiral or capture silk. This type of silk is composed of an
STATEMENT ACCORDING To 37 CPR.
acidi?c glycoprotein, secreted from the ?agelliform gland,
and coated With glue from the aggregate gland, Which makes
A second important type of spider silk is the ?agelliform,
§1.821(c) or (e)iSEQUENCE LISTING
it sticky. The glue is not regarded as silk because it is com
SUBMITTED AS AScII TEXT FILE
posed of glycoproteins and other amino acids. The ?agelli
form silk is exclusively used for the construction of the spiral
Pursuant to 37 CPR. §1.821 (c) or (e), a ?le containing an
ASCII text version of the Sequence Listing has been submit
ted concomitant With this application, the contents of Which
components of the Web. This function results in a ?ber that is
20
believed to alloW for more effective capture of prey.
are hereby incorporated by reference.
Minor ampullate (MI) silk is the spider silk that is secreted
by the minor ampullate glands and is a strong, non-elastic,
deformably stretchable silk used in Web formation (Colgin &
TECHNICAL FIELD
The disclosure relates to silk proteins derived from spider
mite, more speci?cally derived from Telranychus urlicae.
25
mentioned in his representation of the spinning glands asso
ciated to its function that the soft inner silk of the egg sac is
30
produced by the aciniform glands (aciniform silk), Whereas
the tough outer silk of the egg sac is secreted by the cylindrical
or tubuliform spinning glands (tubuliform silk). Viney et al.
(2000) believes the opposite. The tubuliform glands are only
Silk is a secreted, ?brous material that is deposited or spun
by an organism. From a biochemical point of vieW, silk con
sists of protein threads composed of repeating arrays of
polypeptides that contain both discrete crystalline and non
LeWis 1998).
Another spider silk that is discussed in this text is the egg
sac silk that is used to Wrap eggs. Vollrath (1992, 2000)
More speci?cally, the disclosure relates to the use of these
proteins to make ?bers or ?ber-composed material.
BACKGROUND
highly extensible and capable of absorbing the energy of the
?ying prey Without failure. The functional role of the glue is
35
found in female spiders, Which makes it more probable that
the inner silk is indeed secreted by the tubuliform glands.
Because of its attractive properties (high strength, ?exible
crystalline domains that are oriented around a ?ber axis.
With good Water-absorbing poWer, soft, good elastic recovery
Several arthropods, such as spiders, caterpillars mites,
mantids, moths, and beetles, produce silk, or silk-like ?bers.
behavior, glossiness, etc.), silk has a Wide variety of uses in
the apparel, drapery, upholstery and military ?elds. Natural
Insects, as a group, as Well as spiders, produce many different
types of silks and ?brous proteins, such as ?broins and
spidroins. An individual spider may produce as many as nine
different types of silks and ?brous proteins, each of Which
may be composed of more than one type of protein (Kovoor
1987; Haupt & Kovoor 1993). Different silks differ in number
as Well as in sequence of composing proteins. Although all
40
silk has a long history of use as a textile ?ber, and has been
used in recent years for medical sutures, blood vessels, arti
?cial skin, tendons and for binding enZymes (Bunning et al.
1994; KuZuhara et al. 1987). Interest inAnlheraea pernyi silk
for biomedical applications has recently groWn because A.
pernyi SF contains the tripeptide sequence of arg-gly-asp
repeat structures are species dependent and the amino acid
(RGD), knoWn as cell adhesive site for mammalian cell cul
ture (Minoura et al. 1995; Pierschbacher & Ruoslahti 1984a,
1984b; Li et al. 2003). Therefore, it has been investigated as a
composition, as Well as the mechanical characteristics, may
potential biomaterial such as a matrix for the enZyme immo
45
?broin and spidroin proteins do comprise several repeats, the
vary strongly from silk to silk (Zurovec and Sehnal 2002;
Fedic et al. 2003).
Although the domesticated silkWorm Bombyx mori is the
mainstay of the silk industry, there is a considerable trade in
some countries in silk produced by silkWorms living “Wild.”
biliZation and mammalian ?broblast cell culture (KWeon et
50
As each silk has its oWn composition and characteristics,
The most important of these Wild silks are those that are
knoWn as Tussah. Tussah is the product of several species of
silkWorm of the genus Anlheraea, particularly Anlheraea
al. 2001a, 2001b). Silk of the spider Nephila clavipes has
been used to help mammalian neural regeneration (Allmeling
et al. 2006).
there is a lot of interest in the identi?cation of neW silk
55
proteins, opening the possibility for neW applications. Sur
prisingly, We found that spider mites, and particularly Tel
ranychus urlicae, are making silk proteins of Which the amino
acid composition differs rather strongly from that of classical
?broins and spidroins, especially in the alanine, glycine and
60
silks are called Wild, because these Worms are not capable of
serine content. Those differences are found in the global
protein composition, as Well as in the composition of the
being domesticated and arti?cially cultivated. Some
repeats.
mylilla, indigenous to India, and Anlheraea pernyi, Which is
native to China (Huber 1947; Cook 1984). Although Tussah
silk is the most important Wild silk in commercial use, there
are still other varieties of caterpillars that produce silk. These
examples are: Anlheraea yamamai, Allacus ricini, and Ana
DISCLOSURE
cus Atlas.
In recent years, spider silk Was receiving more and more
interest, mainly due to the excellent mechanical characteris
tics of this silk. For spiders, one species can make different
65
A ?rst aspect of the disclosure is a spider mite silk protein,
comprising a sequence selected from the group consisting of
US 8,501,913 B2
3
4
SEQ ID N011 through SEQ ID N0119, or a homologue
“Arti?cially produced,” as used here, means that the ?ber
and/or the composing proteins are not produced by a naturally
thereof. “Homologues,” as used herein, means protein With at
least 70%, preferably at least 80%, even more preferably at
occurring Telranychus urlicae.
least 90% identities, as measured using BLASTp (Altschul et
al. 1997). Preferably, the spider mite is Telranychus urlicae.
Preferably, the proteins have a composition comprising at
5
DETAILED DESCRIPTION OF THE INVENTION
least 40%, preferably at least 45%, even more preferably at
EXAMPLES
least 50% serine and glycine (taking both amino acids
together), Whereby the individual composition of serine and
glycine for each is at least 15%, preferably at least 18%, even
Example 1
more preferably at least 20%, calculated as percentage of the
number of the speci?c amino acid on the total number of
amino acids. Even more preferably, independent of the per
Sequencing of the Telranychus urlicae Genome
centage of glycine, serine is present in at least 21%, prefer
ably at least 26%, even more preferably at least 30%. Even
5
more preferably, the proteins comprise, beside the serine and
glycine content, also at least 15%, preferably at least 17%,
mass produced on the bean plants in groWth chambers at 270
C. and 1618 photoperiod. Plants Were Washed in 0.1% TRI
TON X detergent solution in 2-liter beakers to release all
even more preferably at least 20% of alanine. One preferred
embodiment is a spider mite silk protein, Whereby the protein
is selected from the group consisting of SEQ ID N011, SEQ
ID N013, SEQ ID N014, SEQ ID N018, SEQ ID N019, SEQ
ID N0111, SEQ ID N0112, SEQ ID N0113, SEQ ID N0114,
SEQ ID N0115, SEQ ID N01 16, and SEQ ID N0117.Aneven
spider mite life stages.Adult spider mites, nymphs, larvae and
20
eggs Were ?ltered through series of ?ne sieves to isolate pure
egg fraction. Eggs Were collected in the Eppendorf tube,
treated With bleach solution (to remove plant tissue and pos
sible microbial contaminants) and prepared for the DNA
more preferred embodiment is a spider mite silk protein
Whereby the protein is selected from the group consisting of
SEQ ID N018, SEQ ID N0113 and SEQ ID N0115. The most
preferred embodiment is a spider mite silk protein selected
from the group consisting of SEQ ID N013, SEQ ID N019,
SEQ ID N0112, SEQ ID N0114, and SEQ ID N0117.
25
Another aspect hereof is a nucleic acid molecule encoding
a protein according to the invention, or the complement
thereof, or a functional fragment thereof. “Nucleic acid mol
ecule,” as used herein, refers to a polymeric form of nucle
otides of any length, either ribonucleotides or deoxyribo
nucleotides. This term refers only to the primary structure of
the molecule. Thus, this term includes double- and single
stranded DNA, and RNA under the forms knoWn to the person
skilled in the art, such as, but not limited to, genomic DNA,
cDNA, mRNA, antisense RNA and RNAi. It also includes
30
knoWn types of modi?cations, for example, methylation,
40
“caps” substitution of one or more of the naturally occurring
nucleotides With an analog. 0ne preferred embodiment of a
functional fragment is a fragment that can be used as RNAi.
Still another aspect hereof is a recombinant host cell, com
prising a nucleic acid molecule according to the invention. A
“recombinant host cell,” as used here, is a cell that has been
The London population of T urlicae developed from the
isofemale line in London Ontario, folloWing eight back
crosses (to generate maximum homoZygote population) Was
extraction. Embryos Were ground in the glass tissue grinder
and DNA extraction Was performed using QUIAGEN
Blood&cell culture DNA kit (Midi column #13433) accord
ing to manufacturer’s protocol. DNA for Whole genome
sequencing project Was sequenced using Sanger sequencing
protocol at the Joint Genome Institute (USA Department of
Energy), Walnut Creek, Calif.
Example 2
Identi?cation of the Genes
35
From fragments of ?broin genes available in the database,
blastp and tblastn Were run over the proteome and genome of
Telranychus urlicae. The obtained hits Were all checked
manually as due to the highly repeated nature of the sequence
problems occurred With the prediction and even assembly of
the original genomic sequence. About half of the gene models
Were originally Wrongly predicted, involving incorrectly pre
dicted reading frames. The corrections Were iteratively evalu
ated and aligned using MUSCLE, including the existing
45
?broin genes from the public databases and the already found
(and corrected) genes found in Telranychus urlicae.
genetically modi?ed, preferably by the introduction of a
The originally found proteins all had in common a high
nucleic acid according to the invention. The recombinant host
cell of the invention can be any prokaryotic or eukaryotic cell,
including, but not limited to, bacterial cells such as Escheri
chia coli, yeast cells, such as Saccharomyces spp, Pichia spp,
or Kluyveromyces spp, insect cells, plant cells or mammalian
cells. The recombinant host cells can be used to produce large
percentage of G, A and P organiZed in repetitive patterns. This
50
multiple hits returned, six more genes Were retained, based on
similarity of patterns and coverage by Illumina transcript
quantities of the spider mite silk protein according to the
invention. Methods for the production of recombinant silk
particular aspect Was further used to identify more divergent
proteins having similar patterns. To ?nd them, tblastn Was run
again With the loW-complexity ?lters turned off. From the
55
proteins are knoWn to the person skilled in the art and have
been described, as a non-limiting example, in W09116351
reads. All Were manually annotated and added to the already
found genes, as potentially involved in the ?bers. In total,
tWelve genes Were found having a similar repetitive domain.
Example 3
and W09947661, hereby incorporated herein by this refer
ence.
Another aspect of the invention is the use of a spider mite
silk protein, according to the invention, to make a ?ber. Meth
ods to make arti?cial silk ?bers using silk proteins are knoWn
60
Mechanical and antimicrobial characteristics of the spider
mite silk are investigated. Thread thickness and strength are
to the person skilled in the art and have been disclosed, as a
measured using the standard techniques.
non-limited example, in W00153333 and in Teulé et al.
(2009), hereby incorporated herein by this reference.
Still another aspect of the invention is an arti?cially pro
duced ?ber, comprising one or more proteins of the invention.
Analysis of the Spider Mite Silk
65
The FAVIMAT-ROBOT (Textechno) is used to analyZe the
tensile properties. It is a semi-automatic single-strength
tester, Working according to the principle of constant rate of
US 8,501,913 B2
5
6
extension (DIN 51221, DIN 53816, ISO 5079). The instru
N013, SEQ ID NO:14, and SEQ ID NO:17 have been con
?rmed as being part of the silk by MS.
ment is equipped With a balance allowing the mass to be
measured at a high resolution of 0.1 mg. The instrument
includes a ROBOT, Which is a ?ber storage, equipped With a
Example 5
computer-controlled transfer clamp for the transport of the
single ?ber to the testing position of the FAVIMAT. More
Use of the Polymerase Chain Reaction (PCR) to
Con?rm Gene Expression
over, this instrument is equipped With an integrated measur
ing unit for linear density (in dtex:0.1 g/km). This has the
considerable advantage, certainly for natural ?bers, that the
scopic method (ASTM D 1577iBISFA 1985/1989 chapter
T urticae RNA Was extracted using TriZol reagent (Invit
rogen). Samples for PCR Were prepared by reverse transcrib
ing 3 pg of total RNA using Superscript II Reverse Tran
scriptase (Invitrogen). Aliquots of this reaction Were then
F). The ?ber is preloaded at a prede?ned speed. Further on,
used in PCR reactions. Primers for PCR Were designed to
the ?ber is subjected to an electro-acoustic sinusoidal vibra
tion and the resonance frequency is detected With an opto
electronic sensor. The ?ber linear density is calculated from
amplify short (100-200 bp) fragments from the non-repetitive
?neness is determined simultaneously With the tensile prop
erties. The linear density is measured according to the vibro
the resonance condition, i.e., length, preload, and resonance
frequency of the ?ber. Suggesting a uniform mass distribution
and a round cross-section, the linear density can be calculated
20
as follows:
5' and 3' regions of candidate genes predicted mRNA
sequence. PCR Was performed using Taq DNA Polymerase
(Fermentas) according to manufacturer’s recommendations
and ampli?ed fragments Were cloned into pGEM-T Easy
vector (Promega) for sequencing. SEQ ID NO19, SEQ ID
NO:12 and SEQ ID NO:17 Were con?rmed as being
expressed as mRNA by PCR.
References
Allmeling C., A. JokusZies, K. Reimers, S. Kall, and P. M.
25
In this equation, Tt is the linear density in dtex, Fv is the
preload in cN, f is the resonance frequency and L is the test
length in mm.
As spider mite silk is very resistant to degradation, possible
Altschul S. E, T. L. Madden, A. A. Schaffer, J. Zhang, Z.
Zhang, W. Miller, and D. J. Lipman (1997). Gapped
30
antimicrobial activity of the silk is measured by measuring
BLAST and PSI-BLAST: a neW generation of protein data
base search programs. Nucl. Acids. Res. 2513389-3402.
Bunning T. J ., H. Jiang, W. W. Adams, R. L. Crane, B. Farmer,
and D. Kaplan (1994). In: Silk PolymersiMaterials Sci
ence and Biotechnology, D. Kaplan, W. W. Adams, B.
the inhibition circle around the silk on solid medium
Example 4
35
Con?rmation of the Presence of the Proteins in
Spider Mite Silk by Mass Spectrometry (MS)
Analysis
Ten T urticae adults Were placed into capped and Para?lm
Vogt (2006). Use of spider silk ?bers as an innovative
material in a biocompatible nerve conduit. .1. Cell. Mol.
Med. 10:770-777.
Farmer, and C. Viney (Eds.), American Chemical Society,
Washington DC, ACS Symposium Series, 5441353-358.
Colgin M. A. and R. V. LeWis (1998). Spider minor ampullate
silk proteins contain neW repetitive sequences and highly
conserved non-silk-like “spacer regions.” Protein Science
40
71667-672.
sealed 35 mm Petri plates for 24 hours at room temperature.
Cook J. G. (1984). Handbook of Textile FibresiNatural
Petri plate cap Was removed and examined for signs of mites,
Fibres, MerroW Publishing Co. Ltd., Durham, England,
144-165.
eggs and debris, Which Were removed as necessary. After this,
a cap Was Washed With 1 mL of 95% ethanol and silk threads
suspended in ethanol Were collected in Eppendorf tubes. Con
45
tent of 10-15 tubes Was pooled together and silk threads Were
transferred to a glass container for a Wash With acetic acid.
Silk threads Were transferred back into 95% ethanol, pulled
Huber C. J. (1947). The silk ?bers in Matthew ’s textile
apart, and transferred into Eppendorf tubes With 95% ethanol
for storage and subsequent analysis. Silk thread suspensions
50
Were initially evaporated using a SpeedVac system. The dried
samples Were re-suspended in 75% TFA (tri?uoroacetic acid)
55
injected on a Q-ToF MS system using a 150 minute 0-40%
ACN gradient acquiring data in a data-dependent fashion.
Data analysis Was performed using Peaks Studio 5.2 soft
Ware. Peptides Were matched against T urticae proteome
database. Analysis Was performed both With and Without
Kovoor J. (1987). Comparative structure and histochemistry
of silk-producing organs in arachnids. In: The Ecophysiol
ogy of Spiders, W. NentWig and S. Heimer S. (Eds.),
Springer-Verlag, NeW York, 160-186.
KuZuhara A., T. Asakura, R. Tomoda and T. Matsunaga T.
(1987). Use of silk ?broin for enZymemembrane. J. Bio
60
technol. 51199-207.
KWeon H.Y., I. C. Um andY. H. Park (2001a). Structural and
thermal characteristics of Antheraea pernyi silk ?broin/
chitosan blend ?lm. Polymer 4216651-6656.
KWeon H., S. O. Woo andY. H. Park (2001b). Effect of heat
65
treatment on the structural and conformational changes of
consideration for possible variable post-translational modi?
cations, such as deamidation and oxidation.
Protein ID matches from T urticae proteome database that
appeared in both types of analysis and Were also predicted
using computational approach Were considered for subse
quent ampli?cation and cloning by means of PCR. SEQ ID
?bersiTheir physical, microscopical and chemical prop
erties, H. R. Mauersberger (Ed.), 5th edition, John Wiley &
Sons Inc., NeW York, Chapter XVII, 679-729.
Fedic R., M. Zurovec, and F. Sehnal (2003). Correlation
betWeen ?broin amino acid sequence and physical silk
properties. .1. Biol. Chem. 278135255-35264.
in glass vials. Vials Were then microWaved for 45 minutes at
full poWer in a beaker ?lled With Water. The contents of the
vials Were then dried using a SpeedVac system and, folloWing
this, reconstituted in 10% formic acid. Samples Were then
Haupt J. and J. Kovoor (1993). Silk-gland system and silk
production inMesothelae (Araneae).Annales des Sciences
Naturelles, Zoology, Paris 14135-48.
regenerated Antheraea pernyi silk ?broin ?lms. J. Appl.
Polym. Sci. 8112271-2276.
US 8,501,913 B2
7
8
Li M. Z., W. Tao, S. Kuga andY Nishiyama (2003). Control
ling molecular conformation of regenerated Wild silk
?broin by aqueous ethanol treatment. Polymers for
Advanced Technologies 14:694-698.
Teulé E, A. R. Cooper, W. A. Furin, D. Bittencourt, E. L.
Rech, A. Brooks, and R. V. LeWis (2009).A protocol for the
production of recombinant spider silk-like proteins for arti
?cial spinning Nat. Protoc. 4:341-345.
Minoura N., S. Aiba, Y. Gotoh, M. Tsukada and Y. Imai 5 Viney C. (2000). From natural silks to neW polymer ?bers. J.
(1995). Attachment and growth of ?broblast cells on silk
Text. Inst. 91:2-23 Part 3 Sp. lss. Sl.
?broin. Biochem. Biophys. Res. Commun. 208151 1-516.
Vollrath
F. (1992). Spider Webs and Silks. Scienti?c Ameri
Pierschbacher M. D. and E. Ruoslahti (1984a). Cell attach
can 266152-58.
ment activity of ?bronectin can be duplicated by small
synthetic fragments of the molecule? Nature 309130-33. 10 Vollrath F. (2000). Strength and function of spiders’ silks.
Reviews in Molecular Biotechnology 74:67-83.
Pierschbacher M. D. and E. Ruoslahti (1 984b). Variants of the
cell recognition site of ?bronectin that retain attachment
Zurovec M. and F. Sehnal (2002). Unique molecular archi
promoting activity. Proc. Natl. Acad. Sci. USA 81:5985
tecture of silk ?broin in the Waxmoth, Galleria mellonella.
J. Biol. Chem. 277:22639-22647.
5988.
SEQUENCE LISTING
NUMBER OF SEQ ID NOS:
19
SEQ ID NO 1
LENGTH: 1172
TYPE: PRT
ORGANISM: Tetranychus urt icae
<400> SEQUENCE: 1
Met Asn Ser
1
Lys
Ala Val Gln
Leu Leu Thr Leu
Cys
Leu Val Ile Thr Ala Leu Thr
Thr Asn Ala Asn Ser Leu Phe
Gly
25
Lys
Ile
Gly
Leu
Gly
35
Gly
Asn Met Leu
40
Lys
Pro Phe
Gly
Leu Pro His Met
30
Ile
Asp Gly
Asn
Gly
45
Ser
Lys
Ser Ala Ser Ala Ser Thr Ser Lys Ala Thr Ser
55
60
His His Thr
Gly
Ala
Asn
Gly
Ser
50
Ser Ser Pro Pro Ser
Gly
65
Ala Ser
Gly
Ser Ser
Gly
Ser
85
Gly
90
Pro Pro Ser Leu
80
Ser Ser Ser Ser Ala Thr
95
Asp
Val
Gly
Pro Asn
Lys
Pro Ile Asn Ser His Gly Ser Asn
105
110
Pro Ser Ser
115
Gly
Gln Glu Ser
Gly
120
Ser Ser Ser Asn Ile Ser
125
Tyr
Ala
Asp
Ser Ser
140
Lys
Gln Leu Asn
Ser Thr Thr Ser Ser
150
Gly
Ala Ala Ser
Gly
Ser
Ser Ser
Ser
Gly
130
Ser Ser Thr Ser Ser
135
Ser His
145
Gly
Gly
Ser
Ser
160
Gly
Ser Ala Ala Ser
Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser
180
185
Asp
Ser Ser Ser
190
Ser Ser
165
Gly
Gly
155
Asn
Ser Ala Ala Ser
170
Ser
Gly
Trp
Gly
Pro Thr Thr Ser Thr Ser Thr Ser Asn Ser Pro Asn Ser Ala Ser Ser
195
200
205
Gly
Ser
210
Gly
Ser
Gly
Gly
Ser
Gly
Ser Ser
225
Gly
Ser Ser Ala Ala Ala Ser Ser
215
220
Gly
Ala Ala Ser
Gly
Ser
Gly
Ser
Gly
Ser
Gly
230
Ser
Gly
Ser Ser
245
Gly
Gly
235
Ser
Gly
Ser Ser
250
Gly
Ser Ser
240
Ser Ala Ala Ser
255
Gly
Ser Ala Ala Ser Ser Ser Ala Ala Ala Ala Ala Ala Ser Asp Ser Ser
260
265
270
US 8,501,913 B2
10
—cont inued
Ser Ala Pro Ala Pro Ala Ser Asn Thr
275
280
Gly
Ser
Gly
Ser Ser Ser Ala
285
Ala Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser
290
Ser
305
Gly
295
His Gln Pro Val Asn His Leu Ile Gln His
Pro Thr Ser
Asp
Gln Val
Arg
Gly
320
Ile Ile Ser Ser Ser Phe Ile
325
Ser Ala Ser Ser Ala Ser Gly Ser
340
345
Ser Ala Pro Ala Pro Ala Ser Asn Thr
375
Gly
Gln Ala Ser
Gly
Ser
Gly
Ser
420
Gly
Ser
Ala Ser
Gly
Gly
Ser
Gly
Ser Ser
435
Gly
Ser Ser
Ser Ser Ser Ala
Asp
Ser Ser
400
Gly
Ser Ser Ala Ala Ala Ser Ser
425
430
Gly
Ala
Gly
Ser
Gly
Ser
Gly
Ser
Gly
440
Ser
Gly
Ser Ser
455
Gly
Gly
445
Ser
Gly
Ser Ser
460
Gly
Ser Ala Ala
Ser Ala Ala Ser Ser Ser Ala Ala Ala Ala Ala Ala Ser Asp
470
480
Ser Ser Ser Ala Pro Ala Pro Pro Ser Asn Thr
485
490
Gly
Ser
Gly
Ser Ser
495
Ser Ala Ala Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser
500
505
Ser Ser Ser
515
Gly
Gly
Gly
Gly
Ala Ala Ser
Ser Ser
Ser
Gly
Ser
Gly
535
545
Gly
Asp
Pro Thr Thr Ser Thr Ser Thr Ser Asn Ser Pro Asn
520
525
Ser Ala Ser Ser
530
Ser
Ser
Pro Thr Thr Ser Thr Ser Thr Ser Asn Ser Pro Asn Ser Ala
405
410
415
Gly
Ser
465
Asp
395
Ser Ser
Ser Ser
450
Gly
380
390
Gly
Ser Ser
335
350
Ala Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser
385
Arg
330
Ser Ala Ala Ser Ser Ser Ala Ala Ala Ala Ala Ala Ser
355
360
365
Ser
Ser Ser
315
Gln
Ser
Asp
300
Gly
Ser
550
Gly
Ser
Gly
Ser Ser
Ser
Gly
Ser Ser
585
Ser Ser
Ser Ser Ala Ala Ala Ser Ser
540
Gly
Ser
555
Gly
Gly
Ser
Gly
Ser
Gly
Gln
Gly
Gly
Ser
Gly
Ser Ser
590
Gly
Ser
Ser
Gly
Gln
Gly
560
565
Ser
Gly
Ser Ser
580
Ala Ala Ser
Gly
595
Asp
Ser Ser Ser
610
Gly
Ser Ala Ala Ser Ser Ser Ala Ala Ala Ala Ala Ser
600
605
Gly
Pro Thr Thr Ser Thr Ser Thr Ser Asn Ser Pro
615
620
Asn Ser Ala Ser Ser Gly Ser
625
630
Ser
Gly
Ala Ala Ser
Gly
Ser
Gly
Ser
Gly
Ser Ser
650
Gly
Ser Ser
665
Gly
645
Gly
Ser
Gly
Ser Ser
660
Ser Ala Ala Ser
Ala Ser
Asp
Gly
Gly
Ser
Gly
Gly
Ser Ser Ala Ala Ala Ser
635
640
Ser
Gly
Ser
Gly
Gln
655
Ser
Gly
Ser Ser
670
Gly
Ser Ala Ala Ser Ser Ser Ala Ala Ala Ala Ala
680
685
Ser Ser Ser Ala Pro Ala Pro Ala Ser Asn Thr
Gly
Ser
US 8,501,913 B2
11
—cont inued
690
695
700
Gly Ser Ser Ser Ala Ala Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala
705
710
715
720
Ala Ser Asp Ser Ser Ser Gly Pro Thr Thr Ser Thr Ser Thr Ser Asn
725
730
735
Ser Pro Asn Ser Ala Ser Ser Gly Ser Gly Ser Asp Ser Ser Ser Ala
740
745
750
Pro Ala Pro Ala Ser Asn Thr Gly Ser Gly Ser Ser Ser Ala Ala Ser
755
760
765
Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser Asp Ser Ser Ser Gly
770
775
780
Pro Thr Thr Ser Thr Ser Thr Ser Asn Ser Pro Asn Ser Ala Ser Ser
785
790
795
800
Gly Ser Gly Ser Gly Ser Ser Ala Ala Ala Ser Ser Gly Ala Ala Ser
805
810
815
Gly Ser Gly Ser Ser Gly Ser Gly Ser Gly Gln Gly Ser Gly Ser Ser
820
825
830
Gly Ser Gly Ser Ser Gly Ser Gly Ser Ser Gly Ser Ala Ala Ser Gly
835
840
845
Ser Ala Ala Ser Ser Ser Ala Ala Ala Ala Ala Ala Ser Asp Ser Ser
850
855
860
Ser Ala Leu Ser Pro Ala Ser Asn Thr Gly Ser Gly Ser Ser Ser Ala
865
870
875
880
Ala Ser Asp Ser Ser Ser Gly His Thr Thr Ser Thr Ser Thr Thr Ala
885
890
895
Ser Gly Ser Gly Ser Ser Gly Ser Gly Ser Gly Gln Gly Ser Gly Ser
900
905
910
Ser Gly Ser Gly Ser Ser Gly Ser Gly Ser Ser Gly Ser Ala Ala Ser
915
920
925
Gly Ser Ala Ala Ser Ser Ser Ala Ala Ala Ala Ala Ala Ser Asp Ser
930
935
940
Ser Ser Ala Pro Ala Pro Ala Ser Asn Thr Gly Ser Gly Ser Ser Ser
945
950
955
960
Ala Ala Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser Asp Ser
965
970
975
Ser Ser Gly Pro Thr Thr Ser Thr Ser Thr Ser Asn Ser Pro Asn Ser
980
985
990
Ala Ser Ser Gly Ser Gly Ser Gly
995
Ala Ala
1010
Ser Gly
1025
Ser Ala
1040
Ser Ser Ala Ala Ala
1000
Ser Gly Ser Gly Ser
1015
Ser Ser Gly Ser Gly
1030
Ala Ser Gly Ser Ala
1045
Ser Ser Gly
1005
Ser Gly Ser Gly Ser
Gly Gln Gly
1020
Ser Ser Gly Ser Gly
Ser Ser Gly
1035
Ala Ser Ser Ser Ala
Ala Ala Ala
1050
Ala Ala Ser Asp Ser Ser Ser Ala Pro Ala Pro Ala
Ser Asn Thr
1055
1060
1065
Gly Ser
1070
Ser Ala
1085
Ser Thr
1100
Gly Ser Ser Ser Ala
1075
Ala Ala Ser Asp Ser
1090
Ser Asn Ser Pro Asn
1105
Ala Ser Ser Ser Ala
Ala Ala Ser
1080
Ser Ser Gly Pro Thr
Thr Ser Thr
1095
Ser Ala Ser Ser Gly
1110
Ser Gly Ser
US 8,501,913 B2
14
13
—cont inued
Gly
Ser
Ser Ala Ala Ala Ala Ser Gly Ala Ala Ser
1115
1120
1125
Ser Ser
1130
Gly
Gly Ser Gly Ser Gly
Gln Gly Ser Gly Ser
1135
Gly Ser Gly
Ser Gly Ser
1140
Ser
Ser Gly Ser Ala Ala Ser Gly Ser Thr Val Pro Ala Tyr
1145
1150
1155
Leu Lys
Tyr Thr Asn Glu Ser
1160
1165
Gly Lys Thr Cys Val
Cys Tyr
1170
<210> SEQ ID NO 2
<211> LENGTH: 311
<212> TYPE: PRT
<213> ORGANISM: Tetranychus urticae
<220> FEATURE:
<221> NAME/KEY: miscifeature
<222> LOCATION: (311) . . (311)
<223> OTHER INFORMATION: Xaa can be any
naturally occurring
amino acid
<400> SEQUENCE: 2
Met Arg Thr Leu Gln Ile Leu Leu Val Leu Leu Glu Ile Leu Asp
1
5
10
15
Tyr
Ala Gln Ser Ala Ser Phe Asp Asp Val Ala Leu Gln Ile Asp Pro
Gly
20
25
30
Asn Trp Leu Ile Glu Ala Thr Leu Tyr
35
40
Asp Gln Ser Asn Asp Glu Arg
Tyr Ser Met Arg Glu Met Ile Tyr Ser
Asn Tyr Ser Ile Ser Gly
50
45
Lys
55
Leu Ala Ile Thr Ser Asp Ala Gly Ser Phe Asp
65
'70
75
Ile Phe Tyr His Asp
80
Arg Leu Gln Glu Tyr Arg Leu Val Ile His Asp Asn Arg Cys Asp
Thr
85
Phe Thr Tyr Lys Ser Lys Trp Asp Ser Asn Leu Ser Gly Ile Thr Asn
100
105
Pro Leu Leu Asn Arg Ile Leu Leu Val
115
120
Leu
Asn Trp Gly Gly His Arg Trp Ile
130
135
Arg Gly Thr Thr Met His Ser Asp Tyr
145
150
110
Gly
Pro Ser Leu Ile His
125
Arg
Ser Asp Ser Asp Val Gln Ile
140
Ala Asn Met Asn Gly Asn
155
Lys
160
Leu Arg Val Thr Arg Tyr Phe Lys Ser Lys Glu Ala Ile Gln Pro
165
170
175
Asp
Arg Ile Val Phe Tyr Gly Thr Asp Val
Lys
180
Thr Glu Phe Ser Ser Lys
185
190
Gln Ser Phe Ile Met Asp Phe Thr Ser Val Thr Lys Leu Glu Asn Glu
195
200
205
Val Ser Gly Leu Val Thr Val Thr Pro
210
215
Gly Ile Gly Cys Arg Phe Tyr
220
Leu Glu Ser Ser Ser Pro Ile Pro Asn Val Pro Ser Asn Gln Leu His
225
230
235
240
Tyr Leu Leu Asp Glu Asn Val Lys Gly
245
Pro Arg Pro Ser Ser Lys
250
255
Arg
Glu Glu Val Tyr Ala Asp Ile Glu Ala Gln Leu Leu Tyr Arg Lys Met
260
265
270
Thr Ser Gln Gly Lys Glu Glu Glu Thr Ile Tyr Asp Phe Ser Leu
275
280
Ile Ser Tyr Lys Leu Leu Asp Lys Gly
290
295
Gly
285
Tyr Cys Thr Ile Asp Pro
300
Met
US 8,501,913 B2
15
16
—cont inued
Ala Ser Ser Asp Pro Gly Xaa
305
310
<211> LENGTH: 1112
<2l2> TYPE: PRT
<2l3> ORGANISM: Tetranychus urticae
<400> SEQUENCE: 3
Met Val Phe
1
Thr Asn
Tyr
Lys
Met
Tyr
Val Ser Thr
Leu Asn Leu Leu Ile Leu Ala Ile Thr Ala
10
15
Arg
20
Leu
Asp
Val Asn Met Pro Met
Ser Met
25
Asp
35
Ser Ala Phe Ala
Gly
Ser Asn Ala
55
Gly
Ser Met Pro
Gly
Met Glu
30
Met Met Ser Asn Val Leu
45
Gly Gly
Gly
Ser Glu
Asp
Ala Ala Ser Asn Ala Glu Ser Thr Ala
Thr Glu Asn Glu
Gly
Ala Asn Ala Glu Ala Thr
65
Thr
80
Tyr
Glu Glu Pro
Asp Gly
Glu
Asp Asp Gly
Leu Thr
Tyr Gly
85
Asp
Glu Ser
Asp
Ala
Asp
Ala
Lys
100
Ala
Gly
Asn Asn
130
Ser
115
Asp
Asn
Gly
Ser
Gly
Asn Asn
Gly Gly
Asp
Asn
Gly
Ser
155
Ser Ser Ser Ser Ala Ser
180
Gly
Ser Ser Thr Ser Glu
185
210
Asn Asn Asn
205
Gly
Asn Asn
Ser Ser Ser Ala Thr Ser Ser Ser Ser Ala Ser
215
220
Gly
Ser Ser
Gly
Asn
Gly
Asn Asn
Gly Gly
Gly
Ser
Asp
Asn
Asn
200
Gly
Gly Tyr
Ser
Asn Asn Asn
Ser Ser Ala Ser
260
Gly
Asn Asn
Gly Gly
Asn Asn
275
Gly
Gly Gly
Asn
Gly Tyr Gly
Ser
Gly
305
Asn Asn
Gly Gly
355
240
Asn Asn
Ser
Asp
Gly Gly
Asn
270
Gly
Gly
Ser
Gly
Gly Gly
Gly
Ser
Asn Asn Asn
Gly
Ser Ser Thr Ser Glu
300
Asn
Gly Tyr
315
Asp
Ser
Ser Ser Ser
320
Ser Ser Ser Ala Thr Ser Ser Ser Ser Ala Ser
325
330
335
Ser Ser Thr Ser Glu
340
Gly Gly
Asn Asn
310
Gly
285
295
Gly
Gly
280
290
Asn
Asn Asn Ala
235
Asn
Ser Ser Ser Ala Thr Ser Ser
250
255
Ser Ser Thr Ser Glu
265
Ala Thr Ser Ser Ser Ser Ala Ser
Asn
Gly
230
245
Asp
Ser Ser Ser Ala Thr
Asp
225
Gly
Gly Gly
Ser
190
Thr Ser Glu
Tyr
Asn Asn
Gly
Gly
195
Gly Gly
Gly Tyr Gly
160
Asn Asn
Ser
Gly Gly
Gly
Gly
Gly
Asn
125
Ser Ser Ser Ala Thr Ser Ser Ser Ser Ala Ser
135
140
Asn Asn Asn
165
Asn
Lys
105
Ser Ser Thr Ser Glu Gly Ser
145
150
Gly Tyr
Ala Thr Ala Glu Ser Ala Ala
120
Gly Gly
Asn
95
Asn
345
Gly Tyr
Asn Asn Asn
350
Gly
Gly
Asn
Ser Ser Ser Ala Thr Ser Ser Ser Ser Ala Ser Ser Ser
360
365
US 8,501,913 B2
17
18
—cont inued
Ser Thr Ser Glu
370
Gly Tyr
385
Gly
Ser
Asp
Asn
Gly
Ser
Gly
375
Asn Asn Asn Val Asn Asn
390
Ser Ser Ser Ala Ser
405
Gly
Gly Gly
Asn Asn
380
Asn
Ser Ser Ser Ala Thr Ser
395
400
Arg Asp
Ser Ser Asn
Gly Gly
Leu Thr Met Val
Ala Val Thr Thr Glu Glu Thr Val Ile Thr Thr Met Val Thr Met Glu
420
Asp
425
Gln Ala Gln Gln His His
435
Gln
Arg Asp
450
440
Arg
465
470
Gly
Asn Asn
485
Glu
Gly Gly
Asn Asn
Gly
Gly
Ser Ser Ala Ala Ala Ala Ser
530
535
Gly Gly
Asn Asn
550
Gly
Ser Ser Ala Ala Ala Ala Ser
565
Asn Asn
Gly Gly
Asn Asn
Gln His His His His
Gln Ala
Asn Asn
Gly
490
515
Asn Asn
545
Asp
475
Ser Ser Ala Ala Ala Ala Ser
500
Asn Asn
Pro Gln His Gln Val His Gln Leu
445
Leu Thr Thr Val Ala Val Thr Thr Glu Glu Thr Val Ile
455
460
Thr Thr Met Val Thr
Gln His Ser
Arg
430
Gly
580
Gly
Ser Ser Ser Ala Ala Ala
495
Ser Ser Ala Ser Asn Gly Ser
505
510
Asn Asn
520
Gly
480
Gly
Ser Ser Ser Ala Ala Ala
525
Ala Ser Ala Ser Asn
Gly Gly
Asn Asn
Gly
Ser Ser Ala Ala Ala Ala Ser
625
630
Asn Asn
Gly Gly
Asn Asn
645
Gly
Asn Asn
Gly
Gly
Asn Asn
Gly Gly
Asn Asn
Gly
675
Ser Ser Ala Ala Ala Ser Ser
690
695
Asn Asn
705
Gly Gly
Asn Asn
710
Gly
Ser Ser Ala Ala Ala Ala Ser
725
Asn Asn
Gly Gly
Asn Asn
585
Gly
770
Asn Asn
Gly
Gly
Gly
Ser
575
Ser Ser Ser Ala Ala Ala
Gly
Ser
Gly
Ser
635
Gly
Gly
Gly
Ser Ser Ser Ala Ala Ala
655
Gly
Asp
Ser Ser Ser Ala Ala Ala
685
Ser Ser Ala Ser Asn
700
Asn Asn
Gly
Gly
Asp
640
Ser Ser Ala Ser His Gly Ser
665
670
Asn Asn
680
Asp
Ser Ser Ser Ala Ala Ala
620
Ala Ser Ala Ser Asn
Asn Asn
Asp
Gly
Ser
Asp
Ser Ser Ser Ala Ala Ala
715
720
Ser Ser Ala Ser Asn
730
Gly
Ser
735
Asp
Arg
Ser Ser Ser Ala Ala Ala Ala Ser
745
Gly
Gly
Ser
Asp
Gly Gly
Gly
Asn
Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ala Ser
775
780
Gly
Ser Ser Ala Ser Asn
755
Gly
Asp
His Asn
740
Asn
Ser
Ser Ser Ser Ala Ala Ala
555
560
Ser Ser Ala Ser His
570
650
Ser Ser Ala Ala Ala Ala Ser
660
Gly
540
Ser Ser Ala Ala Ala Ser Ser Asp Ala Ser Ala Ser Asn
595
600
605
Asn Asn
610
Asp
Ser Ser Ala Ser His
Asn Asn
760
Gly
Ser
Asp
Thr Asn
765
Asn Asn
Gly Gly
Asn Asn
Gly
Asn
US 8,501,913 B2
19
—cont inued
785
790
795
800
Asn Gly Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ala Ser Gly
805
810
815
Ser Ser Ala Ser Asn Gly Ser Asp Asn Asn Gly Gly Asn Asn Gly Asn
820
825
830
Asn Gly Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser Ser Gly
835
840
845
Ser Ser Ala Ser Asn Gly Ser Asp Asn Asn Gly Gly Asn Asn Gly Asn
850
855
860
Asn Gly Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ala Ser Gly
865
870
875
880
Ser Ser Ala Ser Asn Gly Ser Asp Asn Asn Gly Gly Asn Asn Gly Asn
885
890
895
Asn Gly Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ala Ser Gly
900
905
910
Ser Ser Ala Ser Asn Gly Ser Asp Asn Asn Gly Gly Asn Asn Gly Asn
915
920
925
Asn Gly Ser Ser Ser Ala Ala Ala Ser Ser Ala Ala Ala Ser Ser Gly
930
935
940
Ser Ser Ala Ser Asn Gly Ser Asp Asn Asn Gly Gly Asn Asn Gly Asn
945
950
955
960
Asn Gly Ser Ser Ser Ala Ala Ala Ala Ser Ala Ala Ala Ala Ser Gly
965
970
975
Ser Asn Ala Lys Lys Asn Asn Gly Ser Asn Asn Ser Gly Ser Asn Ser
980
985
Ala Ala Thr Ser Ser Asn Ser Ser
995
Ser Ser
Ser Gly Ser Ala Ala
Asn Asn Gly Gly Ser
Gly Asn
Ala Thr Ser Ala Ala
Ser Lys Lys Gly Ala
Gly Ser
Lys Gly Ser Asn Gly
Ala Ser Ala Ser Ala
Ala Ala Ser Gly Ala
Lys Ser Gly Lys Ser
Ala Gly Asn
1050
Lys Gln Gly Asn Gly
Pro Gly Asn
1065
Ala Ala Ser Ser Ala
1075
1085
Ser Ala Ala
1035
1060
1070
Arg Gly Asn
1020
1045
1055
Ser Ala
Gly Ser Gly Ser Asn
1030
1040
Asn Ser Gly
1005
1015
1025
Ser Ser
Gly Lys Lys Val Asn
1000
1010
Gly Gln
990
Ser Gly Lys
1080
Pro Ala Lys Gln Gly
1090
Ile Ile Pro
1095
Ala Met Met Ser Lys Ile Pro Thr Leu Ser Val Ser Met Phe
1100
1105
1110
<210> SEQ ID NO 4
<211> LENGTH: 1026
<212> TYPE: PRT
<213> ORGANISM: Tetranychus urticae
<220> FEATURE:
<221> NAME/KEY: miscifeature
<222>
LOCATION:
(972) . . (1005)
<223> OTHER INFORMATION: Xaa can be any naturally occurring amino acid
<400> SEQUENCE: 4
Met Ile Thr Asn Leu Val Phe Leu Cys Leu Phe Leu Thr Thr Cys Ser
1
5
10
15
Leu Ile His Ser Ser His Ser Asn Ser Leu Ser Lys Trp Asn Pro Met
20
25
30
US 8,501,913 B2
21
22
—cont inued
Lys
Ala Ala Ile Ser Ile Pro Met
35
Gly Lys
50
Gly
Met Leu
Asp Gly
40
Ile His Asn Val Thr
Pro
65
Lys
55
Ser Ser
Gly Gly
Ser Ser
Gly
Ser Ala Ala Ala Ala Ser
Lys
Asp
80
Gly
Pro Thr Ser Asn
85
Gly
Gly
Ser Ser
Gly
Ser Ser Ala Ala Ser Ser
105
Gly
Ser
Gly
Ser
100
Gly
Lys His
Pro His Thr Thr Ala Thr Thr Ser
60
Ser Ser Ser
Ser
Glu
45
Asn Ser Ala Asn Ser Ala Ser
90
Ala Ser
Gly
Ser Ser Ser Ser
125
Ala Ala Ala Ser Ala Ser Ser Ser Ala Ala Ser
Asp
Ser
Gly
Gly
Ser Ala Ala
160
Ser Ser
115
Ser
Gly
Gly
Gln
Ser
Gly
Gly
120
130
135
Ala Pro Ala Thr Ser Ser Thr Asn
145
Gly
Ser
150
Gly
Ser
155
Ser Ser Ser Ala Ala Ser Ala Ser Ala Ala Ala Ala Ser
165
170
Ser
Gly
Pro Thr Ser Asn
180
Ser
Gly
Ser Ser
195
Gly
Gly
Asp
Ser Ser
Gly
Ser
225
Ser Ser Ala Ala Ser Ser
200
Gly
Ser
Gly
Gln
Gly
230
Ala Ser
Gly
Gly
Ala Ser
Gly
Ser
260
Gly
Ser
Gly
290
Gly
Gly
Thr Thr Thr Ser Thr Ser Ser Thr Ser
Gly
325
330
Ser
Gly
Gly
Ser
Gly
Ser
Gly
Gln
Gly
340
Gly
Gly
Ser
Gly
Ser
Gly
375
Ala Ser Ala Ser Ala Ala Ala Ala Ser
385
Gly Lys
Gly
Thr Ser Thr Ser Ser Thr Ser
Gly
435
Gly
450
Ser
Gln
Asp
Ser Ser Ser
285
Gly
Gly
Gly
Gly
Gly
Ser
Thr Ser Ile
320
Ala Ser
Gly
Ser
Gly
Ser
335
Gly
Ser Ser Ala Pro Ala Thr
365
Ser Ala Ala Ser Ser Ser Ala
380
Asp
Ser Ser Ser
395
Gly
Pro Thr
400
Gly
Gly
Ser Ser
415
Ser
Ala Ser
Gly
Thr Ser Thr Thr Thr
425
440
Gly
Ser Ser Ala Pro
255
410
Ser Ser Ala Ala Ser Ser
420
Ser
Ser
Ser Ala Ala Ser Ser
Ala Ser
Thr Ala Asn Ser Ala Ser Ser
405
Gly
Gly
315
390
Ser Asn
Thr
Ser Ser Ser Ser Ala Ala Ala Ser
345
350
Ser Ser Ser Ala Ala Ser Asp Ser
355
360
Ser Ser Thr Asn
370
Gly
Asn Ser Ala Asn Ser Ala Ser Ser
295
300
Ser Ser Ala Ala Ser Ser
310
Ser Ser
305
Gly
205
265
Ser Ala Ala Ser Ala Ser Ala Ala Ala Ala Ser
280
Gln Thr Ser Asn
Gly
Ser Ser Ser Ser Ala Ala
235
240
Ser Ser Ser Ala Ala Ser Asp Ser
245
250
Ala Thr Ser Ser Thr Asn
Ser Ser
175
Asn Ser Ala Asn Ser Ala Ser Ser
185
190
Ser Thr Thr Thr Thr Ser Thr Ser Ala Thr Ser Gly Ala Ser
210
215
220
Gly
Ser Ser
140
Ala Ser
430
Gly
Ser
Gly
Ser Ser
Gly
445
Ser Ser Ser Ser Ser Ala Ser
455
460
Gly
Pro His
US 8,501,913 B2
24
23
—cont inued
Ala Asn Ser Met Gln Ser Ile Ala Ser
465
Arg Tyr
470
Ser Ser Ser
Gly
Val Asn Ser Ala
475
Pro Thr Ser Asn
485
480
Asn Ser Ala Asn Ser Ala Ser
490
495
Ser
Gly
Ser Ser Ala Ala Ser Ser
505
Gly
Ala Ser
Gly
Thr Ser Thr Thr Thr Thr Ser Thr Ser Ser Thr Ser
Gly
Ala Ser
Ser Glu Ser Ser
500
Gly
Gly
Asp
515
Gly
Ser
530
Gly
520
Ser Ser
Ala Ala Ala Ser
Gly
545
Gly
Ser
535
Gly
525
Ser
Gly
Gln
Gly
Ser Ser Ser Ser
540
Ser Ser Ser Ala Ala Ser
550
555
Ala Pro Ala Thr Pro Ser Thr Asn
Gly
565
Ser
570
Gly
Asp
Ser
Gly
Ser
Gly
Ser Ala Ala
575
Ser Ser
560
Ser Ser Ser Ala Ala Ser Ala Ser Ala Ala Ala Ala Ser Asp Ser Ser
580
585
590
Ser
Gly
Trp Gly
Pro Thr Ser Asn
595
Ser Ser
Gly
Gly
Asn Ser Ala Asn Ser Ala Ser Ser
600
605
Ser Ser Ala Ala Ser Ser Gly Ala Ser
615
620
Ser Thr Thr Thr Thr Ser Thr Ser Ala Thr Ser
625
630
635
Gly
Gly
Ser
640
Gly
Ser Ala Ala Ser Ser
665
Gly
Ala Ser
Gly
Thr Ser
Thr Thr Thr Thr Ser Thr Ser Ser Thr Ser
Gly
Ala Ser
Gly
Ser
Ala Ser
Gly
Ser
660
650
675
Ser Ser
690
Ser
705
Gly
Gly
Gln
Gly
Thr
Gly
Ser
Gly
Ala Ser
Gly
Ser
645
Ser Ser
Gly
Gly
Gly
680
Gly
Gly
Ser Ser Ser Ser Ala Ala Ala
700
Ser Ser Ser Ala Ala Ser
Asp
Ser
Gly
Ser
Gly
Ser Ala Ala Pro Ser Ser
735
Ser
Gly
695
Gly
Ser
Gly
725
730
Gly
Pro
Asn Ser Ala Asn Ser Ala Ser Ser Gly Ser
760
765
Gly
Ser
740
Thr Ser Asn
Gly
755
Gly
Thr Thr Ser Thr Ser Ser Thr Ser
Gly
785
Gly
Ala Ser
Thr Ser Thr Thr
Ala Ser
Gly
Ser
795
Gly
Ser Ser
800
Gln
Gly
Ser Ser Ser Ser Ala Ala Ala Ser
810
815
Ser Ser Ser Ala Ala Ser
820
Gly
Ser
Ser
Ser
Gly
Gly
780
790
Gly
Asp
745
Ser Ser Ala Ala Ser Ser
775
770
Ser Ser Ala Pro Ala
720
Ser Ser Ser
750
Ala Ala Ser Ala Ser Ala Ala Ala Ala Ser
Gly
805
Ser Thr Asn
835
Gly
Ser
Gly
Gly
Gly
825
Ser
Gly
840
Gly
Ser Ser Ala Pro Ala Thr Ser
830
Ser Ala Ala Ser Ser Ser Ala Ala
845
Ser Ser Ser
860
Gly
Pro Thr Ser
Asn Ser Ala Asn Ser Ala Ser Ser Gly Ser
870
875
Gly
Ser Ser
Ser Ala Ser Ala Ala Ala Ala Ser
850
855
Asn
865
Gly
685
Gln
Ser
Thr Ser Ser Thr Asn
Ser
Ser Ser Ser Ser Ala Ala
655
Ser Ser Ala Ala Ser Ser
Gly
Asp
Ala Ser
Gly
Gly
880
Thr Ser Thr Thr Thr Thr
US 8,501,913 B2
25
26
—cont inued
885
890
Ser Thr Ser Ala Thr Ser
900
Gly
Ala Ser Gly Ser Gly Ser Ser Gly Ser
905
910
Gln
Gly
Ser Ser Ser Ser Ala Ala Thr Ser
920
925
Ser Ala Ala Ser
930
Asp
Ser
Ser
Gly
Gly
895
915
Met
945
Asp
Gln
Asp
Gly
935
Gln
Asp
Ser Ser
Ser Ser Ala Pro Ala Thr Ser Ser Leu
940
Gln Gln Leu Gln Val Gln Gln His Gln His
950
Gln Pro Pro Ala Ala Ser
Cys
955
Arg
960
Phe Ile Ile Arg Xaa Xaa Xaa Xaa Xaa
965
9'70
975
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
980
985
990
Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa
Xaa Xaa Xaa Xaa Xaa Arg Ser Pro
995
1000
1005
Ile Pro
Asn Leu Phe Trp Asp
1010
Gly
Ile
Leu Val Gly Phe Pro
1015
Leu Phe Ile
1020
Phe
1025
SEQ ID NO 5
LENGTH: 2271
TYPE: PRT
ORGANISM: Tetranychus urti cae
SEQUENCE: 5
Met Phe Lys Leu Thr Leu Val Leu Ile Cys Ile Ser Ala Ile Thr Val
1
5
Ser Glu
Gly Arg
10
Ala Val Gln
15
Lys Arg Asn Val Leu Asp Asp Leu Leu
20
Ala Asn Val Gln Ala Thr Ile
35
Leu Pro Ala Val
50
Gly
Gln Ile Glu
Tyr
Tyr
Asp
Ser Val Val
Leu Pro
85
Lys
Asp
Glu Ile
Arg Ala Arg
Ala Leu
Leu Gln
Lys Ile Leu Ser Gln Arg Val Asp
Asp Lys His
Lys Lys
Leu Val Gln Asn Ser Arg
120
Arg
Trp Gly
Phe Glu Asn Leu Ile Ser Met Pro Asp Val
135
140
Ser Val
Gly Thr Ile Asp Lys Ile Glu
155
Lys Ser Arg Gly Leu Phe
185
Lys
Val
Gly Asp
195
Gly Cys Gly
160
Lys Asp Thr Val Val Asp Arg Gly Thr
180
Ala Ile Ala Glu
190
Thr Ile Arg Cys Phe Phe Gly Ile
200
Asn
Arg Lys
Lys Thr Leu
125
Leu Ile Ser Glu Val Ser Ser Leu
165
170
Ile Asn
Lys
110
Phe Met Leu Gln Asn Asp Gly
Ser Thr Glu Ser Ile Phe Gln Ile Leu
145
150
Gly
80
105
Ser Leu Glu Ser
130
Lys His
Ser Ser Phe Leu Glu
60
75
115
Arg
Lys Leu Ser
45
Ala Ala Ala Gln Gln Gly Thr Asp Asn Phe
100
Ile
Val Glu Asp Asp Gly
55
Ser Ile Ser Gln Leu
65
Glu Ser
Lys
40
210
Ser
215
Lys Arg
Gln Glu Glu Glu Glu
Lys
205
Ala
Asp Glu Glu Lys Asp Gln Lys
220
Lys Lys Arg Lys Gln Gln Glu Lys Ile
US 8,501,913 B2
27
28
—cont inued
225
Glu
230
Lys
Glu
Lys
235
Glu Asn Val
Arg
Ala Ala Leu Glu
245
Ala
Asp Arg
Asp Lys
Asp
Ser Asn Ser Pro Thr
295
Ser Ser
305
Lys
Thr Asn Thr
Lys
Lys
Lys
310
Thr Phe
Asp
Asp
Asn Ser
Lys Lys
Gln Ser Ser
Gly
355
Asp
Pro Ser Ala Ala
360
Leu Ser Pro Gln
370
Val Val
Asp
385
Ser Asn
Lys
Lys
Asn
Gly
Lys Lys
Ser
Gly
Glu Ser Glu Ala Ser
Asp
Ala Ser Asn Glu
Thr Ser Ser Pro Gln
Lys
350
Pro Val
Lys Lys
470
Lys Lys Asp
Asp
Asp
Gly
440
Ser Glu Ser Ser Ser
445
Ser Glu Ser Ser Ser Thr
475
Lys
Lys Lys Asp
Lys
Gly
Ala Gln
Gly
Gly
Gln Thr Ala
480
495
Val Glu Ser Ser
Lys
Gln
510
Lys
Ser Ser Pro
Gln Ser
525
Leu Val Val
Asp
Ala Pro Val Ser Thr
540
Thr Ser Asn
Asp Lys Lys
Ser Ser
555
Pro Ser Ala Thr
565
Lys
Asp
560
Thr Ala Glu Ser
Lys Lys
575
Val Gln Leu Val Val
Asp
Leu Pro Val Ser Ser
585
Gly
Lys
630
Thr Ser Asn Asn
Lys Lys
600
Ser Ser Glu Pro Ser Ser Thr
610
615
Lys Asp
Thr
400
Gln Ser Glu Pro Ser Ser
535
Gly
Lys
505
Lys Lys Asp
Val
Ser Gln Asn Thr Asn Ser
595
Ser Glu Ser Ser
625
Gly
Ala Val Glu Ser
490
Thr Asn Ser
Ser Ser Ser
580
Leu Thr
Val Gln Leu
Lys
Ser
550
Ser Thr Ser
Gly
Lys
520
Ser Ser Ser
530
Lys Lys
Gln Thr Ala Glu Pro
Ala Ser Ser Gln
Ser Gln Ala Thr Glu Ser
Asp
Ser
Lys
Lys
Ser Ala Gln Asn Ala Glu Pro
500
Ser Leu
545
Asp
Gln Ser
365
485
Asp
Thr
335
Leu Pro Val Ser Thr Ser Ser Asn Thr Asn Ser
455
460
465
Glu Pro
Asp Lys
345
Lys Lys
Asn Thr
320
425
Ser Ser Pro Gln
435
Leu Ile Val
450
Pro
Lys
Pro Ser Thr
410
420
Lys Gly
Lys
Ser Glu Ser Ser Ser
380
405
Lys
Ser Ser Ala Ser Ser
300
Leu Pro Val Ser Ala Ser Gln Asn Thr Asn Ser
390
395
Asp Lys Lys
Gln Ala
Val Ile
Pro Thr Ser Asn Ser Glu
315
340
Arg
285
Ser Gln Ser Gln Ala Gln Pro Val Thr Leu
325
330
Glu Thr
Lys
270
Ala Glu Asn Ser Ala
Lys
290
Val Glu
255
280
Glu Val
Lys
250
Leu Ser Asn Glu Glu Lys Ile
260
265
Ser Ala Ala Leu
240
Lys
Gln Thr Val Glu Pro Thr
Lys Gly
620
Ser Glu Ala Ser Thr
635
Ser Ala Pro Gln
645
Ser Glu Pro
605
Lys
Lys
Gln Asn
Asp
640
Gln Ser Asn Ser Ser Ser
650
Glu
655
Gly
US 8,501,913 B2
29
—cont inued
Val Gln Leu Val Val Asp Thr Pro Val Ser Asn Ser Gln Asp Thr Arg
660
665
670
Ser Gly Asn Ser Asn Asp Lys Lys Ser Ser Asp Ser Ser Ser Glu Pro
675
680
685
Ser Ser Thr Lys Gln Thr Val Glu Pro Thr Lys Asp Ser Glu Ser Ser
690
695
700
Lys Gln Ser Gln Ala Ser Glu Ser Lys Lys Asp Ser Ser Ser Gly Val
705
710
715
720
Gln Leu Val Val Asp Thr Pro Val Ser Ser Gly Ser Ser Asp Arg Asn
725
730
735
Gln Pro Thr Asp Thr Lys Lys Asp Val Asp Ser Ser Glu Lys Thr His
740
745
750
Asn Ser Glu Ser Lys Ile Asn Glu His Glu Thr Ser Thr Lys His Ser
755
760
765
Asp Leu Tyr Ser Gln Thr Val Thr Gln Ala Trp Asn Ala Glu Ser Leu
770
775
780
Ser Ala Gly Gln Asp His Thr Thr Lys Pro Asn Ala Ser Leu Ser Asp
785
790
795
800
Glu Thr Ala Val Glu Phe Ser Ser Asp Ser Tyr Glu Asp Val Thr Val
805
810
815
Gly Ser Ala Ala Ser Ser Glu Thr Ser Asn His Gly Ser Ile Ser Val
820
825
830
Ala Ala Thr Ser Glu Ala Asn Gln Pro Thr Thr Gln Ser Thr Asn Ser
835
840
845
Ser Thr Ser Asp Gly Asn Lys Val Val Val Ile Ile Thr Ser Asn Asp
850
855
860
Asn Asp Ser Gly Ser Ser Glu Ile Pro Ser Gln Ser Ser Asn Gln Gln
865
870
875
880
Thr Ser Ser Asn Ser Ala Ser Ala Thr Asn Asn Gln Thr Ser Gln Glu
885
890
895
Ser Ser Ser Thr Ile Thr Ser Val His Asp Gly Val Asn Ala Gly Ser
900
905
910
Asp Gln Ala Lys Asp Gln Ser Gly Ser Pro Ser Ser Gln Thr Ser Asn
915
920
925
His Glu Ser Ser Leu Ser Ser Thr Ser Glu Ser Thr Thr Gln Ser Ser
930
935
940
Gln Ala Ser Tyr Glu Ser Ser Ser Ile Arg Thr Ser Asp Thr Glu Ser
945
950
955
960
Asn Ser Pro Val Thr Gln Gln Ser Gly Gly Leu Ser Ile Asp Val Thr
965
970
975
Val Gly Ser Ile Val Pro Val Ser Thr Glu Thr Lys Cys Arg Asn Arg
980
985
Asp Pro Gln Met Lys Asn Asp Ser
995
Glu Ser
1010
Gly Ser
1025
Ser Ser
1040
Glu Ile
1055
Gln Ser
1070
990
Ala Ser Ser Val Gln
1000
Asn Ser Thr Val Ala
1015
Ala Val Thr Glu Asn
1030
Leu Glu Tyr Ser Thr
1045
Arg Thr Ser Asp Ser
1060
Ser Glu Val Ser Ile
1075
Ala Ser Gln
1005
Ser Leu Tyr Val Asp
Ser Thr Val
1020
Gln Ser Val Ser Gln
Thr Ser Thr
1035
Gln Ala Ser Ser Gln
Glu Ser Gly
1050
Glu Ser Ser Asn Pro
Leu Ser Gln
1065
Asp Val Thr Val Gly
1080
Ser Val Asp
US 8,501,913 B2
31
32
—cont inued
Ser Val
Ala Thr Glu Thr Ser
1085
1090
Gln Ala Ser Gln
Gln Ser
Ser Val Ser Ile
Thr Ser Glu
1110
1100
Gly
Ser Ser Asn Val Ser
1105
Asn Glu Pro Thr Thr Ser Asn Thr Ser
1115
1120
Asn Thr Val
1125
Gly
Asp
Phe
Ser Ser
Gln Ser Ser Ser His Gln
1145
1150
Asp
Val Ser Ser
Val Thr Thr Asn
1130
Asp
Asp Gly
Asn
Val Ile
Thr Ser Ser
1095
1135
Ser
Ala
1140
Gly
Thr
Leu Asn Glu
1155
Pro
Thr Glu Ser Gln Asp Leu Thr Thr Thr Ser Glu Ser Leu
1160
1165
1170
Ser
Asp
Glu Ile
Asp
Phe Glu
1175
Val Thr
Phe Ser Thr
Val
Gly
1190
1220
Val Val
Thr
1250
Val Ser Val Ser
Tyr
Val Val Val Ala Thr
Glu Glu
Tyr
Glu
Ala Thr Thr
1215
Asn Glu Pro Thr Thr
Ser Asn Ser Ala Asp
1225
1230
1235
Gly
Ser
1185
Ser Ser Ala
Ser Ser Ala Thr Ser
Asn
1195
1200
Ser Gln
Ser Ser Asn His Glu
1205
1210
Gln Ser
Asp
1180
Asn Gln Asn
Asp
1240
Gly
Asn Thr
Ser
Ala Ser Ser
1245
Pro Ser Gln Ser Ala Asn Gln Gln Thr Ser
Ser Ser Ser
1255
1260
Ser Ser Ala Thr Asn Ser Gln Ala Ser Gln Glu Ser
Asn Pro Thr
1265
1270
1275
Val Ala
Ser Leu
Tyr
Glu
1280
Glu Asn
Asp
Ser Ile Val
Gln Ser Val Ser Gln
1295
Ser
Ala Val Thr
1290
Thr Ser Thr Ser Ser
1300
Ser Glu
Tyr
1305
Ser Thr
Gln Ala Ser Ser Gln Glu Ser
1310
1315
Asp
Gly
1285
Gly
Ala
Lys
Arg
Thr Ser
1320
Ser
Glu Ser Ser Asn Pro Val Ser Gln Gln Ser
Ser Glu Val
1325
1330
1335
Gly
Ser Ala
1340
Asp
Thr Ser
Ser Gln Ala Val Gln
Thr Ser Ser Gln Ser
Ser Ser Asn
1360
1365
1355
Val Ser
Val Thr Val
1345
Ala Ser Val Ser Ser
1370
1400
Lys
Asp Gly
Asn Thr Val Val Val Ile Val
Ser Ser Asn
1390
1395
Asp Asp
Ser Val
Ser Gln Ser Val Thr
Pro Ala
1420
1425
Asn Ser Ala Ser Leu His Glu
1430
1435
Leu Ser
Asp
Glu Ile
1445
Tyr
Glu
1450
Asp
Ala Thr Val
1460
Val Ser
Thr Thr Ser
1380
Ser Glu Val Ser Ser
Ser Gln Ser Ala Ser
His Glu Ser
1405
1410
Pro
Ser
1415
Glu Ser
Glu Val Asn Glu Pro
1375
Ser Thr
Ser
1385
Glu Asn
Ser Ile Val Pro Val
Ser Thr Glu
1350
Thr Thr Ser Glu
Gly
Gly
Gln
Asp
Trp
Leu Thr
Thr Ser Ser
1440
Ala Phe Glu Phe Ser
Thr
Asp
Ser
1455
1465
Ser Ser Ala Ser Ser
Ser Ile Ser
1470
Gly
Asn Glu Pro Thr Thr
Gln Ala Thr
US 8,501,913 B2
33
34
—cont inued
1475
1480
Ser Ser
Thr Ser
1490
1485
Asp Gly
Asn
Thr Val Val Val Ile Val Thr Thr
1495
1500
Gly
Ser
Ser Ser Ala Pro Ser
Gln Thr Ser
1510
1515
Asn Glu Asn Glu Ser
1505
Ser Gln Gln Thr Asn Ser Gly
Ser Ala Ala Asn Asn
Gln Ala Ser
1520
1525
1530
Gln Glu
1535
Glu Val
Ser Asn Pro Thr Val
Ala Ser Asn Phe Asp
Ser Ile Ser
1540
1545
Gln Ser Thr Gln Ser
1550
Thr Thr
Val
Gly
1565
Met Pro
Thr Gln
1580
Ser Asn
Ser
1595
Gly Gly
Ala Ser Ser Ser
1555
Asp
Tyr Asp Asp
1560
Ser Ser Glu Ala Val Val Gln Ser
Thr His
1570
1575
Asp
Lys
Ser Thr
Pro Ser Ser Ser Thr
Gln Ser Thr
1585
1590
Ser Glu Ser
Ser Asn Pro Val Thr
Gln Gln Ser
1600
1605
Val Ser Ile
Asp
1610
Val
Thr Val
Gly
1615
Ser Val
1620
Asp
Ser Val
Ser Thr
Glu Thr Ser Ser Gln Ala Ser Gln Thr Ser
Ser Gln Ser
1625
1630
1635
Thr Ser
1640
Ala Val
Asn Thr Ala Asn Ser Ala Ala
1645
Val Val Phe Val Thr
1655
Phe
Gly
Gly
Thr Glu Ala Thr
1660
Ile Pro Ser Gln Ser
1670
Thr Ser Ser Ser Ser
1675
Gln
Asp
Thr Val
Gly
Trp
Phe Glu Phe Ser Thr
Asp
Thr
Gly
Ser
Ser Ser Ser
Asp
Lys Tyr
Glu Ser Ser
1695
Asn Ser
Gly
Ser Leu Ala
1710
Asn Thr Asn Ala Ser
1720
1730
Ala
1680
Ser Ser
Glu Thr Ile Thr Gln Ala
1700
1705
1715
Gly
1665
Ser Glu
Ile Asn Asn Gln Ser Ser Glu Gln
1685
1690
Val Glu
Ser Ser
1650
Ser
Gly Gly
Leu
Ser
1725
Tyr
Asp
1735
Glu
Asp Gly
Gln Thr Val
1740
Ser Val Val Thr Gln Asp
Gln Ser Val Ser Pro
Thr Ser Ser
1745
1750
1755
Ser Ser
Ser Glu
1760
Tyr
Ser Thr
Gln Ser Ser Gln Ser
Ser Gln Gln
1765
1770
Ser Glu
Ser Thr
1775
Arg
Asn Ser Asn Ser Glu Pro Ser
Asn Pro Val
1780
1785
Thr Gln
Asp
Glu Val
Gln Ser
1790
Val Ser
1805
Ser Asn
1820
Ser Ile
Thr Glu Ser Gln
Asp
1810
1850
Val Thr
1855
Gly
Ser
Pro Gln Thr Ser Ser
Gln Ser Ser
1815
Ser Ile Ser Val
Ser Thr Ser Ser Glu
1825
1830
Ile Val Thr Thr Asn
Val
1800
Pro Thr
Thr Gln Ala Thr Ser Ser
1835
1840
Val Val
Asp
1795
Asp
Lys
Ser
Asp Gly
Gly
Asn Glu
Asn Thr Val
1845
Ser Ala Ser Ser
1860
Gly
Thr Pro
Ser Gln
Thr Ser Asn Gln Gln Thr Ser Ser Ser Ser
Ser Ser Val
1865
1870
1875
US 8,501,913 B2
35
36
—cont inued
Ile Asn
Asn Gln Ala Ser Gln Glu Asn
1880
1885
Thr Phe
Asp
Phe Ile Ser Glu
1895
Gln
1925
Val
Tyr Asp Asp
Thr
Ser Ser Ser
1905
Thr Val
Gly
1915
Glu Ser
Tyr
1955
Asp
Ala Pro Ser Thr Gln
Ser Ser
Glu Val Val
1920
Ser Thr Ser Gln Thr
Ser Ser Gln Asn Thr
Thr Pro Ser
1930
1935
Ser Ser
1940
Gln Ser
Pro Thr
Val Ala Ser
1890
1900
Ser Pro
Ser
1910
Asp
Arg
Gly
Ser
Ile
1945
Arg
Asn Ser
Thr Val
Gly
Ser Val
1975
Asp
Ser
Asp
Ser
1950
Ser Pro Val Thr
Gln Gln Ser
1960
1970
Asp
Gly
Glu
Val Ser Ile
1965
Ser Val Ser Thr
Glu Thr Ser
1980
Ser Gln
Ser Thr Gln Thr Ser
Ser Gln Ser Ser Ser
Asn Val Ser
1985
1990
1995
Val Ser
Ser Thr Ala Ser
Glu
2005
2000
Ala Ser
Ser
2015
Ser Ser Ser Ala Gln
2020
Gly
Ser Ile
Arg
Thr Ser
2030
Thr Gln
Gln Ser Ser Ala Ile
Val
Asp
2060
Asn Glu Pro
Thr Thr Ser
2010
Gly
Thr Gln Ser
Ser Gln Glu
2025
Asp
Ser Glu Ser Ser
Asn Pro Val
2040
Asp
Ile
2035
2045
Gly
Asp
Val Thr
2050
Val
Gly
Val
Asp
Ala Val Val
2090
Val Phe Val Thr Ser
2095
Tyr
Thr Thr
Gln Ser
2105
Ala Ser
Ser
Ser Val Ser Ser Glu
Thr Ser Ser Gln Ala
Ser Gln Thr
2065
2070
Ser Ser
Gln Ser Thr Ser Asn Thr Ala Asn Ser Ser Ala
2075
2080
2085
Ser
Gly
2055
Gly
2110
Phe His Phe Thr Asn
2120
Gly
Ser
Thr Glu Ala
2100
Ser Ser Ser Ser Ser
Ser Ser Ser
2115
Gln Thr Ser Gln Val
2125
Asn Glu
Asp
2130
Asn Glu
Pro Ala Val Ser Thr
Glu Thr Ile Gln Val
2135
2140
2145
Asp
Gln Thr
Ser Thr
Gln Ser Ser Ser Gln Glu Ala Val Ser Thr
Ser Ser Ala
2150
2155
2160
Ser Ser
Glu Thr
2165
Lys
Asn Pro
2170
Thr Gln Pro Ala
Val
Asp
Thr
Thr
Arg
Val
2175
Ser Ser
Ser Glu Ser Ser His
Ala Phe
2180
2185
Asp
Glu Ile
2190
Ser Thr
Pro Leu Glu Ser Ile
Thr Glu Ala Val Asn Glu Val Asn
2195
2200
2205
Asn Glu
Ser
2210
Asp
Ser Thr Glu Ala Ser Gln Ile Thr
Ser Thr
2215
2220
Asn Ala
Ser His Asn His Thr
2225
2230
Tyr
Val Ala Val
2255
Leu Ser Ser Ser Ser Asn Ser Ala
2260
Pro Ala Ala
2270
Lys
Val Ser
Lys
Val
2235
Ser Thr
Glu Pro Ile Val Ala Ser Ser Val Ala Lys
2240
2245
2250
Thr Val
Gly
Asp
Val
2265
Lys
Val Val Ile
US 8,501,913 B2
37
38
—cont inued
<2lO> SEQ ID NO 6
<211> LENGTH: 337
<2l2> TYPE: PRT
<2l3> ORGANISM: Tetranychus urticae
<400> SEQUENCE: 6
Met Ile Arg Ala Ala Leu Phe Ile Ala Leu Phe Ala Leu Ala Thr Ala
1
5
10
15
Ala Asn Leu Ser Leu Asp Ser Gln Trp Glu Ser Phe
20
Lys
Ile
Lys Tyr
25
Gly Lys Ser Tyr Glu Ser Glu Ala Glu Glu Thr Tyr Arg Arg Ser Val
35
40
45
Phe Ala Lys Lys Met Glu Lys Ile Lys Ala His Asn Glu Arg Ala Asp
50
55
Asn Gly Glu Val Thr His Arg
65
Lys Gly
Ile Asn
Lys
Phe Ser
Asp
7O
Thr Thr Glu Glu Phe Lys Ala Lys His Leu
85
Gly
Leu Thr Ala
90
Leu
80
Lys His
95
His Gly Ser Arg Ser Ile Val Arg Arg Ser Ala Pro Leu Ile His Asn
100
105
Ala Asn Asn Thr Val Lys Ala Ala Ala
115
11O
Tyr
Val
Asp Trp Arg
120
Gly Ile Val Ser Gln Val Lys Glu Gln Gln Asp Cys
130
Thr
Lys
125
135
Gly
Ala
Cys Trp
140
Ala Phe Ser Ala Ile Ala Ala Ile Glu Ala Ala Asn Ala Gln
145
150
155
Lys
Asp Cys
Ser
175
Gly Lys Leu Val Glu Leu Ser Val Gln Asn Val Leu
165
170
Asn Tyr Ser Ser Leu Gly Cys Ala Gly
180
Gly Trp
Ile Asn
185
Tyr
Thr
160
Trp
Ala Phe
190
Ser Tyr Val Lys Asp Asn Lys Gly Ile Asp Thr Glu Lys Ser Tyr Pro
195
200
205
Tyr Ile Ser Gly Asp Gly Ile Asp Tyr His Thr Cys Arg Tyr Asn Glu
210
215
220
Ser Asn Lys Gly Ala Ser Ile Ala Ser Phe Val
225
230
Asp
Ile Pro Glu
235
Gly
240
Asp Glu Glu Ala Leu Leu Ala Ala Val Ala Glu His Val Val Ala Val
245
250
255
Gly Ile Asp Ala Ala Ser Val Tyr Glu Tyr Glu Ser
260
Gly
265
Ile
Tyr Tyr
270
Thr Asp Glu Cys Ser Ser Asp Pro Lys Asp Asn Asn His Ala Val Ala
275
280
285
Val Val Gly Tyr Gly Ser Glu Asn Gly Ile Pro Phe Trp Ile Ile Lys
290
295
300
Asn Ser Trp Gly Met Leu Phe Gly Glu Ser
Gly Tyr
305
315
310
Phe
Arg
Leu
Tyr
320
Arg Gly Ser Asn Met Cys Gly Ile Ala Asn Gly Ala Ser Tyr Pro Ile
325
Val
<211> LENGTH: 317
<2l2> TYPE: PRT
<2l3> ORGANISM: Tetranychus urticae
330
335