HYPERPRODUCTION OF A BIFUNCTIONAL HYBRID PROTEIN, METAPYROCATECHASE-PROTEIN A, BY GENE FUSION

被引:12
作者
KOBATAKE, E
IKARIYAMA, Y
AIZAWA, M
MIWA, K
KATO, S
机构
[1] TOKYO INST TECHNOL,DEPT BIOENGN,MEGURO KU,OOKAYAMA 2-12-1,TOKYO 152,JAPAN
[2] SAGAMI CHEM RES CTR,GENET ENGN SECT,NISHIOHNUMA,KANAGAWA,JAPAN
关键词
Binding protein; Enzyme conjugate; Fusion protein; Recombinant DNA;
D O I
10.1016/0168-1656(90)90067-L
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
A hybrid protein between metapyrocatechase and Staphylococcal protein A was produced by recombinant DNA techniques. A plasmid carrying the fusion gene that encodes the hybrid protein was constructed and expressed in E. coli. Over 70% of soluble proteins of the cell extracts was estimated to be the hybrid protein. This fusion protein is about 65000. Both the IgG-binding activity of protein A and the metapyrocatechase activity were found in the hybrid protein. The optimum pH of metapyrocatechase in the fusion protein was at around 6.5 and Km was 1.3 × 10-5 M. A simple immuno-enzymometric assay was developed for anti-BSA antibody using the fusion protein. © 1990.
引用
收藏
页码:87 / 96
页数:10
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