Overexpression in Pichia pastoris and crystallization of an elicitor protein secreted by the phytopathogenic fungus, Phytophthora cryptogea

被引：33

作者：

ODonohue, MJ ^{[1
]}

Boissy, G ^{[1
]}

Huet, JC ^{[1
]}

Nespoulous, C ^{[1
]}

Brunie, S ^{[1
]}

Pernollet, JC ^{[1
]}

机构：

[1] INRA, UNITE RECH BIOCHIM & STRUCT PROT, F-78352 JOUY EN JOSAS, FRANCE

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1996年 / 8卷 / 02期

关键词：

D O I：

10.1006/prep.1996.0098

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A synthetic gene encoding beta-cryptogein, a member of the elicitin family, has been cloned into a vector for expression by the methylotrophic yeast, Pichia pastoris. Having first optimized the gene construction for secretion, we have overexpressed a modified beta-cryptogein in a secreted form. A purification scheme suited to this expression system has been developed and highly pure, biologically active protein has been obtained. For structural analysis of this recombinant beta-cryptogein, and new mutated forms thereof, optimal conditions for the crystallization of this protein have been determined and crystals that diffract to 2.2 Angstrom have been obtained. (C) 1996 Academic Press, Inc.

引用

页码：254 / 261

页数：8

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