Utilization of the methoxymalonyl-acyl carrier protein biosynthesis locus for cloning of the tautomycin biosynthetic gene cluster from Streptomyces spiroverticillatus
Utilization of the methoxymalonyl-acyl carrier protein biosynthesis locus for cloning of the tautomycin biosynthetic gene cluster from Streptomyces spiroverticillatus
复制标题
DOI：
10.1128/jb.00172-06
复制
发表时间：
2006-06-01
影响因子：
3.2
通讯作者：
Shen, Ben
中科院分区：
文献类型：
--
作者：
li, Wen Li;Ju, Jianhua;Shen, Ben
他莫霉素（TTM）是一种有效的蛋白磷酸酶抑制剂，由含有螺缩酮部分的聚酮链和带有二烷基马来酸酐结构的酰基链组成。使用简并引物PCR从螺旋轮叶链霉菌中克隆形成甲氧丙二酰-酰基载体蛋白的基因。发现该位点包含5个基因（ttmC、ttmA、ttmD、ItmB和ttmE），其中之一被用作探针以克隆110-kb TTM生物合成基因簇。通过基因失活和突变互补实验证实了ttmA基因参与TTM生物合成。
Tautomycin (TTM), a potent protein phosphatase inhibitor, consists of a polyketide chain containing a spiroketal moiety and an acyl chain bearing a dialkylmaleic anhydride structure. PCR using degenerate primers was used to clone genes from Streptomyces spiroverticillatus for formation of the methoxymalonyl-acyl carrier protein. This locus was found to contain five genes (ttmC, ttmA, ttmD, ItmB, and ttmE), one of which was used as a probe to clone the 110-kb TTM biosynthetic gene cluster. The involvement of the ttmA gene in TTM biosynthesis was confirmed by gene inactivation and mutation complementation experiments.