A kind of bacterial strain that improves the production of tetramycin Z and the method for preparing tetramycin Z
A technology of tetramycin and bacterial strains, applied in the field of microorganisms, can solve the problems of high randomness, time-consuming and laborious, and difficulty in increasing the production of natural products such as antibiotics, and achieve the effect of increasing production
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Embodiment 1
[0027] Example 1: Obtaining of low-concentration streptomycin-resistant mutants (the first round of screening of drug-resistant mutants)
[0028] Prepare a GYM solid plate with a low concentration of streptomycin at a final concentration of 30 μg / mL, that is, the concentration of streptomycin in the GYM solid plate is 30 μg / mL. Use a pipette gun to draw 100 μL of spore suspension of Streptomyces chromogenes D (1 × 10 6 cells / mL) on the GYM solid plate, spread evenly with a sterile applicator stick, culture in an incubator at 28°C for 7 days, pick a single colony to a new GYM solid plate containing 20 μg / mL streptomycin (one single bacterium colony to draw a flat plate), put it in an incubator at 28°C for 5 days, and the bacterial strain that can grow again on the new GYM solid plate is the streptomycin-resistant mutant. A total of 15 low-concentration streptomycin-resistant mutant strains were obtained, numbered fz1-fz15 respectively. The 15 mutant strains were subjected to l...
Embodiment 2
[0029] Embodiment 2: Obtaining of high-concentration streptomycin-resistant mutant strains (the second round of drug-resistant mutant strain screening)
[0030] Prepare a GYM solid plate with a final concentration of streptomycin of 300 μg / mL, that is, the concentration of streptomycin in the GYM solid plate is 300 μg / mL. Use a pipette gun to draw 1000 μL of low-concentration streptomycin-resistant mutant strain fz9 spore suspension (1×10 12 cells / mL) on the GYM solid plate, spread evenly with a sterile applicator, place in an incubator at 28°C for 10-15 days, pick a single colony to a new GYM solid plate containing 300 μg / mL streptomycin (A single colony is drawn on a plate), placed in an incubator at 28° C. for 7 to 10 days, and the bacterial strain that can grow again on the new GYM solid plate is a high-concentration streptomycin-resistant mutant. A total of 3 high-concentration streptomycin-resistant mutant strains were obtained, numbered respectively sz2-1, sz2-2 and sz...
Embodiment 3
[0031] Embodiment 3: Obtaining of rifampicin-resistant mutants (the third round of screening of drug-resistant mutants)
[0032] Prepare a GYM solid plate with a final rifampicin concentration of 20 μg / mL, that is, the rifampicin concentration in the GYM solid plate is 20 μg / mL. Use a pipette to draw 100 μL of the mutant sz2-3 spore suspension obtained in the second round of screening (1×10 10 cells / mL) on the GYM solid plate containing 20 μg / mL rifampicin, spread evenly with a sterile applicator, place in a 28°C incubator for 7 days, and pick single colonies to new ones containing 20 μg / mL rifampicin Fuping's GYM solid plate (one single colony draws one plate), put it in an incubator at 28°C for 5 days, and the bacterial strain that can grow on the new GYM solid plate again is the rifampicin-resistant mutant. In this screening, two rifampicin-resistant mutants were obtained, numbered tz3-1 and tz3-2.
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