黑暗链霉菌中tbmA基因的功能研究  

Functional analysis of tbmA gene of Streptomyces tenebrarius

作　　者：杨钧[1] 夏焕章[1] 余永红[1] 刘晓辉[1] 

机构地区：[1]沈阳药科大学生物制药系,辽宁沈阳110016

出　　处：《微生物学杂志》2007年第4期31-34,共4页Journal of Microbiology

基　　金：沈阳市科学技术计划项目(1063310-1-00)

摘　　要：PCR获得tbmA基因内部863 bp片段,构建基因阻断穿梭载体pSPU112-1,经接合转移导入Strepto-myces tenebrariusH6,筛选单交换阻断变株,并用Southern blot验证阻断变株的tbmA已经被破坏。经发酵产物分析,阻断变株不再合成氨甲酰妥布霉素,只合成安普霉素。首次从分子水平证明了tbmA只参与氨甲酰妥布霉素生物合成,而不参与安普霉素的生物合成。A 863 bp amplification fragment of tbmA gene of Streptomyces tenebrarius was obtained by PCR.A recombinant plasmid pSPU112-1 containing the 863 bp fragment for gene blocking was constructed and then the plasmid was transfered into S.tenebrarius H6 by way of conjugation.The result of southern blot assay indicated that the tbmA gene of the blocked mutant was exactly disrupted.The mutant could synthesize apramycin nut no longer produced carbamoyltobramycin,which is firstly confirmed that tbmA gene is closely related to carbamoyltobramycin biosynthesis but not involved in apramycin biosynthesis.

关 键 词：黑暗链霉菌 氨甲酰妥布霉素 生物合成基因 基因阻断 

分 类 号：Q933[生物学—微生物学] R978[医药卫生—药品]