[Overexpression and detection of the mutated glucose isomerase GIG138P and GIG138P-G247D in Streptomyces lividans].
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.
Department of Molecular and Cell Biology, School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
Guo-Ping
GP
Zhu
Ying
Y
Zhang
Yang
Y
Xu
Jian-Guo
JG
Tang
Chong
C
Xu
D019747:Aldose-Ketose Isomerases / Q000032:analysis / Q000235:genetics*; D015153:Blotting, Western; D005822:Genetic Vectors; D009154:Mutation; D013302:Streptomyces / Q000235:genetics*
D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase