IMMOBILIZATION AND SINGLE-STEP PURIFICATION OF FUSION PROTEINS USING DEAE-CELLULOSE

被引：84

作者：

SANCHEZPUELLES, JM ^{[1
]}

SANZ, JM ^{[1
]}

GARCIA, JL ^{[1
]}

GARCIA, E ^{[1
]}

机构：

[1] CSIC,CTR INVEST BIOL,UNIDAD GENET BACTERIANA,VELAZQUEZ 144,E-28006 MADRID,SPAIN

来源：

EUROPEAN JOURNAL OF BIOCHEMISTRY | 1992年 / 203卷 / 1-2期

关键词：

D O I：

10.1111/j.1432-1033.1992.tb19840.x

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have developed a new single-step system, using a DEAE matrix, to immobilize and/or purify fusion proteins containing the choline-binding domain of the Streptococcus pneumoniae murein hydrolases. We have constructed a choline-binding-domain - beta-galactosidase chimera, which can be purified by this procedure and shows a high beta-galactosidase activity when immobilized in the column. A vector plasmid, pCUZ1, containing the lpp(p)-5/lac promoter as well as 13 restriction sites, was constructed to facilitate the cloning and expression of gene fusions. This plasmid also allows the selection of recombinants by the well-known blue/white 5-bromo-4-chloro-3-indolyl-beta-D-galactoside procedure. A chimera between the choline-binding domain and the pneumococcal hemolysin was also constructed and purified using pCUZ1.

引用

页码：153 / 159

页数：7

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