Overexpression and detection of the mutated glucose isomerase GIG138P and GIG138P-G247D in Streptomyces lividans.
- Author:
Guo-Ping ZHU
1
;
Ying ZHANG
;
Yang XU
;
Jian-Guo TANG
;
Chong XU
Author Information
1. Department of Molecular and Cell Biology, School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
- Publication Type:Journal Article
- MeSH:
Aldose-Ketose Isomerases;
analysis;
genetics;
Blotting, Western;
Genetic Vectors;
Mutation;
Streptomyces;
genetics
- From:
Chinese Journal of Biotechnology
2002;18(3):304-307
- CountryChina
- Language:Chinese
-
Abstract:
The shuttle expression vectors pHZGI1 and pHZGI2 were successfully constructed by inserting structural genes of GI containing single mutated site G138P and double mutated site G138P-G247D into E. coli-Streptomyces shuttle vector pHZ-1272, respectively. Then they were transformed into S. lividans TK54 strain by protoplast transformation. SDS-PAGE indicated that two shuttle vectors in TK54 strain expressed obviously specific bands at 42.5 kD after inducted by 2 micrograms/mL thiostrepton. Optical densitometric scan showed that the content of the mutant enzymes GIG138P and GIG138P-G247D were about 19% and 22% of dissoluble proteins, respectively. Western blotting farther proved that GIG138P and GIG138P-G247D were expressed in S. lividans TK54.
