Increase of the expression of midecamycin 4"-hydroxyl propionyltransferase gene (mpt) by a promoter-like fragment from the midecamycin producing strain.
The promoter region of the midecamycin 4"-hydroxyl propionyltransferase gene (mpt) gene was reconstructed by PCR, and ligated with a fragment from the midecamycin producing strain (Streptomyces mycarofaciens var. 68) which contained fairly strong promoter activity (PLF). Recombinant plasmids pCHFPE2 (promoter region of mpt was reconstructed by combining the PLF in the upstream of its own promoter) and pCHFPE3 (the promoter of mpt was replaced by PLF) were obtained. The extent of expression of mpt was measured according to the amount of propionylspiramycin bioconverted from exogenous spiramycin by transformants of S. lividans TK24 containing pCHFPE2 and pCHFPE3. The results showed that the PLF could increase the expression of mpt in S. lividans TK24 up to 89.02% and 58.53%, respectively, and also enhance the expression in the industrial spiramycin producing strain S. spiramyceticus to a great extent.
Gu
H
H
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences, Beijing, China.
Wang
Y
Y
Xu
X
X
Wei
X
X
Fong
M
M
D000217:Acyltransferases / Q000235:genetics*; D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D017931:DNA Primers; D015870:Gene Expression; D007933:Leucomycins / Q000096:biosynthesis*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism
D000900:Anti-Bacterial Agents; D017931:DNA Primers; D007933:Leucomycins; D000217:Acyltransferases; C078622:midecamycin 4''-propionyltransferase; C026483:midecamycin