Wang Xiang-Jing, Guo Suo-Lian, Guo Wan-Qian, Xi Di, Xiang Wen-Sheng
School of Life Science, Northeast Agricultural University, Harbin, China.
J Antibiot (Tokyo). 2009 Jun;62(6):309-13. doi: 10.1038/ja.2009.33. Epub 2009 May 15.
To investigate the function of nsdA in Streptomyces bingchengensis, it was cloned and sequenced, which presented an 89.89% identity with that of S. coelicolor. The lambdaRED-mediated PCR-targeting technique was used to create nsdA replacement in the S. bingchengensis_226541 chromosome. The nsdA disruption mutant, BC29, was obtained, which produced more pigment and spores than did the ancestral strain. HPLC analysis revealed that the disruption of nsdA efficiently increased milbemycin A(4) production and nanchangmycin production by 1.5-fold and 9-fold, respectively. Complementation of the nsdA mutation restored the phenotype and antibiotic production. These results showed that nsdA negatively affected sporulation and antibiotic production in S. bingchengensis.
为了研究冰城链霉菌中nsdA的功能,对其进行了克隆和测序,结果显示它与天蓝色链霉菌的nsdA具有89.89%的同一性。利用λRED介导的PCR靶向技术在冰城链霉菌226541染色体上实现nsdA替换。获得了nsdA缺失突变体BC29,它比原始菌株产生更多的色素和孢子。高效液相色谱分析表明,nsdA的缺失使米尔贝霉素A(4)产量和南昌霉素产量分别有效提高了1.5倍和9倍。nsdA突变的互补恢复了表型和抗生素产量。这些结果表明,nsdA对冰城链霉菌的孢子形成和抗生素生产具有负面影响。