Optimized production method of abamectin
A technology of abamectin and its production method, applied in the field of biomedicine, can solve problems such as toxicity, no relevant research reports on genes, complex chemical components, etc.
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Embodiment 1
[0038] Embodiment 1 The improvement of Streptomyces avermitilis target antibiotic output
[0039] With the application of molecular biology and bioengineering technology in the field of antibiotic breeding, genetic engineering breeding technology has become the main means of improving industrial production strains. The invention improves the output of antibiotics by improving the active efflux efficiency of antibiotics. The specific operation is to connect the target gene responsible for antibiotic efflux to the carrier in vitro through genetic engineering technology, and then transfer it into recipient cells, so that the antibiotic efflux gene can be efficiently expressed and inherited in the recipient.
[0040] (1) Construction of plasmids (used for high expression of target genes transferred into hosts)
[0041] Plasmid construction for overexpression of sav933 and sav934 genes:
[0042] The F cosmid Fosmid 20B4 of Streptomyces avermitilis NRRL 8165 genomic library was di...
Embodiment 2
[0078] Embodiment 2 prepares the industrial high-yield bacterial strain of Abamectin
[0079] Step 1. Transfer of plasmids into Streptomyces hosts and screening and verification of conjugative transfers
[0080] The high-copy plasmid pFAveT of sav933 and sav934 was transferred into Streptomyces avermitilis industrial strain 3-115 cells by conjugative transfer, through the verification of thiostrepton resistance, the plasmid was extracted from Streptomyces and reversed Escherichia coli DH10B, and screened A correct genetically engineered strain was named 3-115 / pFAveT. Then the empty vector pJTU1278 was transformed into Streptomyces avermitilis 3-115 cells by conjugative transfer, and the control strain 3-115 / pJTU1278 was screened by the same method.
[0081] The above 3-115 / pFAveT, that is, Streptomyces avermitilis 3-115 / pFAveT was submitted on December 15, 2011. ), the General Microorganism Center CGMCC of the China Committee for Culture Collection of Microorganisms, and its...
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