重组TaqDNA聚合酶在大肠杆菌中表达条件的优化  

Optimization of Recombinant Taq DNA Polymerase Expression in E.coli

作　　者：何钢[1] 邢伟一[1] 罗丽华[2] 石慧[1] 韩文军[1] 

机构地区：[1]中南林学院生命科学与技术学院,湖南株洲412006 [2]海南大学,海南海口570000

出　　处：《中南林学院学报》2004年第4期62-65,共4页Journal of Central South Forestry University

摘　　要：　以插入重组TaqDNA聚合酶基因的pET-24a表达质粒转化大肠杆菌菌株,转化菌铺平板的最佳量为50至100μL,可获得转化菌的单菌落,单菌落扩大到200mL培养量,表达TaqDNA聚合酶,经SDS-PAGE分析,表达带的相对分子质量约为82ku,最佳表达条件为接种量1/500,工程菌培养到OD600值为1.0开始诱导,以2mmol/LIPTG诱导8h,TaqDNA聚合酶可获得较高的表达效率.The plasmoid containing recombinant Taq gene was transformed into E.coli strain Bl_(21)(DE)3. Optimal inoculation dosage to flat plate of the translation bacterium is 50～100 μL. The positive clones from the LB media plate with Kan were cultured to OD_(600)=0.6. Induced with 2 mmol IPTG for eight hours, all clones seem to have a 94 KD recombinant Taq protein expression band by SDS-PAGE analyses. It is the best for this protein expression to culture the OD_(600) to 1.0, induced with 2 mmol IPTG for eight hours.

关 键 词：生物化学 重组Taq DNA聚合酶 大肠杆菌 表达条件 

分 类 号：Q784[生物学—分子生物学] R378.21[医药卫生—病原生物学]