庆大霉素生物合成关键酶基因在E.coli中的克隆与表达

Cloning and expression of the key gene of gentamicin biosynthesis in E.coli

从庆大霉素产生菌?棘孢小单孢菌基因组中扩增出参与庆大霉素生物合成的关键酶基因——2-脱氧青蟹肌糖合成酶基因GntB,将其分别克隆到克隆载体pBS-T和表达载体pET-22b(+)上,并将pET-gntB转化大肠杆菌E.coliBL21(DE3),用IPTG诱导使GntB基因实现表达.GntB基因大小为1 193 bp,其编码的氨基酸含397个残基,约为42 kD的多肽链.

The gene GntB, the key gene to participate in gentamicin biosynthesis was cloned from Micromonospora echinospora. Then the GntB gene was cloned into clone vector pBS - T and expression vector pET -22b（ ＋ ）, constructed the expression vector pET -gntB and cloned into E. coli BI221 （DE3）. GntB protein was expressed by IPTG induction in E. coli BI221 （ DE3 ）. The length of GntB gene was 1 193 bp and coded 397 AA（about 42 kD）.