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us
20090226969Al
(19)
UnitedStates
(12)
PatentApplicationPublication
Johanssonetal.
(10)
Pub.No.:
US
2009/0226969
Al
(43)
Pub.Date:
Sep.10,2009
(54)SPIDERSILKPROTEINSANDMETHODSFORPRODUCINGSPIDERSILKPROTEINS(75)Inventors:JanJohansson,Stockholm(SE);GoranHjiilm,Uppsala(SE);MargaretaStark,Hagcrsten(SE);AnnaRising,Uppsala(SE);StefanGrip,Uppsala(SE);WilhelmEngstrom,Saltsjo-Boo(SE);My
Hedhammar,
Stockholm(SE)CorrespondenceAddress:BIRCHSTEWARTKOLASCH
&
BIRCHPOBOX747FALLSCHURCH,VA22040-0747(US)(73)Assignee:SPIBERTECHNOLOGIESAB,Uppsala(SE)(21)Appl.No.:12/087,289(22)PCTFiled:Dec.28,2006(86)PCTNo.:PCT/SE2006/001505§371(c)(I),(2),(4)Date:Jun.30,2008(30)ForeignApplicationPriorityDataDec.30,2005(SE)0502932-7PublicationClassification(51)Int.Cl.
C12P21100C07K141435C07H21100C07K11107
(2006.01)(2006.01)(2006.01)(2006.01)(52)U.S.Cl.435/69.7;530/353;536/23.4;530/402(57)ABSTRACTTheinventionprovidesanisolatedmajorampullatespidroinprotein,whichconsistsoffrom150to420aminoacidresi-duesandisdefinedbytheformulaKEP-CT.KEPisarepeti-tive'N-terminallyderivedproteinfragmenthavingfrom80to300aminoacidresidues.CTisaC-terminallyderivedproteinfragmenthavingfrom70to120aminoacidresidues.Theinventionfurtherprovidesanisolatedfusionproteinconsist-ingofafirstproteinfragment,whichisamajorampullatespidroinprotein,andasecondproteinfragmentcomprisingafusionpartnerandacleavageagentrecognitionsite.Thefirstproteinfragmentiscoupledviasaidcleavageagentrecogni-tionsitetothefusionpartner.Theinventionalsoprovidesamethodofproducingamajorampullatespidroinproteinandpolymersthereof.
PatentApplicationPublicationSep.10,2009Sheet1of8US
2009/0226969
Al
7
4371
107135171198235266294330
357
3944214sa
4895175S3581618648
676
712
740
776
804840
668
904932969
999
10281060
FIG
1
PatentApplicationPublicationSep.10,2009Sheet2of8US
2009/0226969
Al
A
C=====::JAAAAMAAAAAAAA
IZZZZ2I
ASASAAASAASTVANSVS/natorASAASAAAS/hyb
~GQG(G/S)QGG(QlY)GG(L/Q)GQGGYGQGAGSS~GQGGQGQG(G/R)YGQG(AlS)G(S/G)S
::::>
G(R/Q)GQG(G/R)YGQG(AlSN)GGN
...---Nephila-likeGly-richrepeat
+
+
B
++++++++
+
4Gly/AIa
2Gly/Ala-CTaggregates
FIG2
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