伊钮小单孢菌2-脱氧蟹肌醇氨基转移酶基因sisM的克隆与表达  

Cloning and expression of 2-deoxy-scyllo-inosose aminotransferase gene sisM from Micromonospora inyoensis

作　　者：曾志红[1] 洪文荣[1] 罗敏玉[2] 戈梅[2] 陈代杰[3] 

机构地区：[1]福州大学生物科学与工程学院 [2]上海来益生物药物研发中心,上海201203 [3]上海医药工业研究院,上海200040

出　　处：《中国抗生素杂志》2008年第5期269-272,共4页Chinese Journal of Antibiotics

摘　　要：从伊钮小单孢菌(Micromonospora inyoensis)中扩增到参与西索米星生物合成基因sisM,其开放阅读框长1263bp,编码含421个氨基酸残基(44.3ku)的蛋白质。经Blast比对,该蛋白与已报道的L-谷氨酰胺:2-脱氧蟹肌醇(DOI)氨基转移酶有很高的同源性,据此推测sisM是编码DOI氨基转移酶的基因。该基因在IPTG诱导下.在大肠埃希菌BL21(DE3)实现了表达,表达产物的分子量与推测的一致。这是从伊钮小单孢菌克隆到的又一参与西索米星核心基团2-脱氧链霉胺(DOS)生物合成的新基因,该基因在GenBank的登录号为EU074791。The gene sisM, a novel gene involved in sisomicin biosynthesis was cloned from Micromonospora inyoensis by PCR. Its ORF is 1,263 bp coding 421 amino acids （44.3ku）. A Gapped-BLAST search showed that the deduced sisM gene product is highly homologous to the previously reported L-gluatamine. 2- deoxy-scyllo-inosose aminotransferase. Based on protein homologies, the product of sisM is suggested as a L- gluatamine.2-deoxy-scyllo-inosose aminotransferase. Then it was over-expressed by IPTG induction in E. coli BL21 （DE3）. The molecular weight observed in the SDS-PAGE was similar to that calculated from amino acid sequences of sisM. sisM is another DOS biosynthetic gene involved in sisomicin biosynthesis in M. inyoensis. Its accession number in GenBank is EU074791.

关 键 词：sisM L-谷氨酰胺:2-脱氧蟹肌醇氨基转移酶 西索米星 伊钮小单孢菌 

分 类 号：R978.19[医药卫生—药品] R978[医药卫生—药学]