黑暗链霉菌中新脱氢酶基因的克隆与功能初步研究

S.tenebrarius H6主要产生安普霉素、氨甲酰妥布霉素和氨甲酰卡那霉素B,其中后两者仅在3′位有差异。为克隆相关基因,根据已报道的aprD3、livY、gentY基因设计兼并性PCR引物,并采用SON-PCR(singleoligonucleotide nested PCR)方法和LASP-PCR(linear amplification single primer PCR)方法,扩增获得部分SCD1基因,Blast分析为短链脱氢酶基因。采用基因阻断技术,使S.tenebrariusH6中SCD1基因失活。与野生菌株相比,变株的抗生素组分和含量几乎没有变化,但变株产孢子能力下降,且产孢子时间推迟,推测该基因与孢子的生成调节相关。

Cloning and Function of a Novel Dehydrogenase Gene from Streptomyces tenebrarius

Streptomyces tenebrarius（St）H6 mainly produces ampramycin,carbamoyltobramycin and some carbamoylkanamycin B,the only difference between the latter two is in 3＇ position.In order to clone the corresponding genes,according to the reported genes aprD3,livY,gentY,an annexational PCR primer were designed and obtained partial SCD1 gene adopting SON-PCR and LASP-PCR methods,Blast analysis showed that SCD1 was a short chain dehydrogenase gene.Adopting gene obstruction method the SCD1 gene in St H6 was deactivated.There was little remarkable change in the antibiotic production and content between the wild strain and mutants,but the sporulation ability declined and delayed suggesting that SCD1 gene was related to spore formation and regulation.