Overexpression and detection of the mutated glucose isomerase GIG138P and GIG138P-G247D in Streptomyces lividans / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 304-307, 2002.
Article
in Chinese
| WPRIM
| ID: wpr-231329
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Streptomyces
/
Blotting, Western
/
Aldose-Ketose Isomerases
/
Genetic Vectors
/
Genetics
/
Mutation
Type of study:
Diagnostic study
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2002
Type:
Article
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