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

被引：11

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

来源：

JOURNAL OF BIOTECHNOLOGY | 1990年 / 16卷 / 1-2期

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

页码：87 / 96

页数：10

共 18 条

[11] NOZAKI M, 1968, J BIOL CHEM, V243, P2682
[12] NOZAKI M, 1963, BIOCHEM Z, V338, P582
[13] Nozaki M., 1970, METHODS ENZYMOLOGY, V17, P522, DOI 10.1016/0076-6879(71)17235-7
[14] GENETIC CONSTRUCTION AND HIGH-LEVEL GENE-EXPRESSION IN ESCHERICHIA-COLI OF A PRECURSOR OF SALMON CALCITONIN-I
OHMORI, M
NARUSHIMA, H
MIKI, T
IKARI, T
NUMAO, N
KONDO, K
[J]. AGRICULTURAL AND BIOLOGICAL CHEMISTRY, 1988, 52 (11): : 2823 - 2830
[15] ELECTROPHORETIC TRANSFER OF PROTEINS FROM POLYACRYLAMIDE GELS TO NITROCELLULOSE SHEETS - PROCEDURE AND SOME APPLICATIONS
TOWBIN, H
STAEHELIN, T
GORDON, J
[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (09) : 4350 - 4354
[16] UHLEN M, 1984, J BIOL CHEM, V259, P1695
[17] UHLEN M, 1983, GENE, V23, P369, DOI 10.1016/0378-1119(83)90025-2
[18] METABOLISM OF TOLUENE AND XYLENES BY PSEUDOMONAS (PUTIDA (ARVILLA) MT-2 - EVIDENCE FOR A NEW FUNCTION OF TOL PLASMID
WORSEY, MJ
WILLIAMS, PA
[J]. JOURNAL OF BACTERIOLOGY, 1975, 124 (01) : 7 - 13

← 1 2 →