Mutant capable of raising cyclic lipopeptides antibiotic output by using RNA polymerase mutation, its preparation method and its application
An RNA polymerase and mutant technology, which is applied in the field of RNA polymerase mutant screening, can solve problems such as low yield, and achieve the effect of stabilizing physiological and biochemical characteristics and genetic traits
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[0027] 1. Screening of RNA polymerase mutants
[0028] Activation of the strain
[0029] Inoculate wild-type Bacillus CC09 with 1% inoculum in 4 mL / tube of LB liquid medium, activate and culture at 37°C, 120 rpm for 48 hours, and then perform secondary activation in 8 LB liquid cultures with the same inoculum amount and culture conditions culture medium for 24 hours.
[0030] Preparation of rifampicin-containing plates
[0031] Add the rifampicin solution to the LB solid medium at 50-60°C to a final concentration of 50 μg / mL, which is the rifampicin-containing plate.
[0032] Will The secondary activation seed solution (8 test tubes) prepared in , was evenly spread on the plate containing rifampicin according to the amount of 200 μL / dish, cultured in the dark at 37°C for 2 days, and the colonies were observed after 2 days, which was the possible mutation of RNA polymerase A total of 78 mutants were screened.
[0033] 2. Determination of the content of high-yield muta...
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