Abstract： Objective To establish and optimize a genetic manipulation system suitable for Streptomyces fradiae, and to obtain a recombinant strain with increased neomycin titer through molecular modification. This study lays a theoretical foundation for the subsequent acquisition of high-yield strains of neomycin. Methods The optimal spore production medium and resistance selection markers of Streptomyces fradiae were selected, the key influencing factors of the conjugation system were optimized, and a recombinant strain over-expressing the neoC gene was obtained to further study the effect of the neoC gene on the neomycin titer. Results The optimal medium for Streptomyces fradiae conjugation was the A75 medium. The optimal spore treatment conditions were heat-shocked at 50℃ for 10min and precultured at 37℃ for 4h. The optimal ratio was 10:1. The optimal addition time of apramycin was 30℃ for 15h. After optimization, the conjugation frequency increased by 7.69 times. At the same time, we obtaineda neoC gene over-expressing strain SF-neoC with 22.29% increase in neomycin titer. Conclusion The genetic operation system of Streptomyces fradiae was successfully established and optimized, providing a basis for other applications based on the subsequent molecular transformation of Streptomyces fradiae.
孙俊峰 王珊 刘鹏飞 薛正莲* 王洲 刘艳. 费氏链霉菌遗传操作体系的优化与应用[J]. 中国抗生素杂志, 2021, 46(02): 128-133.
Sun Jun-feng, Wang Shan, Liu Peng-fei, Xue Zheng-lian,Wang Zhou and Liu Yan. Optimization and application of genetic manipulation system of Streptomyces fradiae. CJA, 2021, 46(02): 128-133.
