Method for improving streptomyces bambergiensis bambermycin yield and bacterial strain
A technology of Streptomyces canola and flavomycin, which is applied to the field of Streptomyces canola strains to increase the production of flavomycin, can solve problems such as increasing the production of Streptomyces canola, achieve a convenient and quick genetic operation method, and overcome blindness. performance, increase productivity
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[0016] Example: Improving Flavomycin Production by Interrupting the nsdA Gene in a Streptomyces spectae strain
[0017] (1) PCR cloning of a fragment comprising the nsdA gene from the genomic DNA of Streptomyces spectrum
[0018] (a) inoculate Streptomyces spectrum ZM2006 in 30mLTSB culture medium (as described later), cultivate 36hr at 37 ℃, collect mycelium, extract genomic DNA; Template, the universal primer P obtained by bioinformatics analysis 1 : GAAGATCTCATGGACGAGCTGGGCAA and P 2 : GAAGATCTCTTGATCTCGTCGAGCCG PCR amplification, PCR reaction program: 95 ℃ pre-denaturation 5min; 95 ℃ denaturation 45 seconds, 58 ℃ renaturation 1min, 72 ℃ extension 5min, 30 cycles; 72 ℃ extension 5min. Agarose gel electrophoresis detection, a band of about 4.7kb was obtained. (c) carry out agarose gel recovery with the 4.7kb band that (b) obtains, carry out enzyme linkage reaction with carrier pMD18-T (the AT cloning kit purchased from TaKaRa Company); (d) obtain with (c) The enzyme-link...
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