METHOD FOR PRODUCING WEB PROTEIN, A FUSED PROTEIN, RECOMBINANT DNA, AN EXPRESSION VECTOR, A HOST CELL AND STRAIN-PRODUCERS

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EP 2 644 616 A8

(12)	CORRECTED EUROPEAN PATENT APPLICATION
	published in accordance with Art. 153(4) EPC

(15)	Correction information:
	Corrected version no 1 (W1 A1)

(48)	Corrigendum issued on:
	03.09.2014 Bulletin 2014/36

(88)	Date of publication A3:
	02.10.2013 Bulletin 2013/40

(43)	Date of publication:
	02.10.2013 Bulletin 2013/40

(21)	Application number: 10859885.5

(22)	Date of filing: 13.12.2010

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International Patent Classification (IPC):

C07K 1/00^(2006.01)
C07K 14/435^(2006.01)
C12N 15/62^(2006.01)
C12N 15/81^(2006.01)

C07K 19/00^(2006.01)
C12N 15/12^(2006.01)
C12N 15/63^(2006.01)
C12N 1/16^(2006.01)

(86)	International application number:
	PCT/RU2010/000752

(87)	International publication number:
	WO 2012/070977 (31.05.2012 Gazette 2012/22)

(84)	Designated Contracting States:
	AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

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Priority:

25.11.2010 RU 2010148011

(71)	Applicants:
	Bogush, Vladimir Grigorievich Moscow 119607 (RU) Beburov, Mikhail Yurievicha Moscow 119607 (RU) Debabov, Vladimir Georgievich Moscow 115409 (RU)

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Inventors:

KOZLOV, Dmitriy Georgievich
Moscow 117623 (RU)
GUBAYDULLIN, Irek Ilyasovich
Ufa 450059 (RU)
DAVYDOVA, Lyubov Ivanovna
Podolsk 142116 (RU)
ZALUNIN, Igor' Arsenevich
Moscow 117593 (RU)
SIDORUK, Konstantin Vasilevich
Podolsk 142100 (RU)
CHEPEREGIN, Sergey Eduardovich
Moskovskaya obl. 140005 (RU)
BOGUSH, Vladimir Grigorievich
Moscow 109544 (RU)
BEBUROV, Mikhail Yurievich
Moscow 119607 (RU)
DEBABOV, Vladimir Georgievich
Moscow 115409 (RU)

(74)	Representative: Icosa
	83 avenue Denfert-Rochereau 75014 Paris 75014 Paris (FR)

(54)	METHOD FOR PRODUCING WEB PROTEIN, A FUSED PROTEIN, RECOMBINANT DNA, AN EXPRESSION VECTOR, A HOST CELL AND STRAIN-PRODUCERS

(57) The invention relates to the field of biotechnology and provides a method for producing recombinant proteins from the orb-weaving spider silk in yeast cells. This involves the construction of an expression vector which comprises a DNA sequence encoding a recombinant protein of the orb-weaving spider silk fused with a sequence encoding an ubiquitin-like protein occupying an N-terminal position with respect to the spider silk recombinant protein within the fused protein. The expression of a hybrid gene makes it possible to increase tens of times the production of recombinant spider silk protein, wherein the recombinant protein accumulates in the yeast cells in a water-insoluble fraction in the form of a processed protein free of a hybrid component.
The invention also relates to fused proteins comprising sequences of recombinant proteins of the orb-weaving spider silk and of ubiquitin-like proteins, to recombinant DNAs encoding the fused proteins, to host yeast cells and to expression vectors suitable for carrying out the method, and also to producer strains of recombinant proteins of the orb-weaving spider silk.