[Expression of the polyketide ketoreductase gene from midecamycin producing strain in E. coli].
Two primers were designed and synthesized according to the polyketide ketoreductase from midecamycin producing strain gene(MKR) sequence data which has been reported, MKR gene was amplified using PCR technique. The amplified MKR gene was subcloned into NdeI and BamHI sites of the T7RNA polymerase-dependent pT7-7 expression vector, and introduced into E. coli K38/pGP1-2. Proteins were isolated from transformant. The result of exclusive labeling by L-35S-methionine of plasmid-encoded proteins SDS-PAGE analysis showed that the MKR was produced in E. coli. The expressed MKR has bioactivity.
Xia
H
H
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100050.
Wang
Y
Y
D000429:Alcohol Oxidoreductases / Q000096:biosynthesis / Q000235:genetics*; D001426:Bacterial Proteins; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005784:Gene Amplification; D015870:Gene Expression; D007933:Leucomycins / Q000096:biosynthesis*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis
D001426:Bacterial Proteins; D007933:Leucomycins; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; C087192:polyketide synthase ketoreductase; C026483:midecamycin