兔IFRG基因的克隆及表达分析  

Cloning and expression analysis of rabbit IFRG gene

作　　者：彭丽婵[1] 丁彪[2] 钱怀剑[1] 李文雍[3] 

机构地区：[1]安徽大学生命科学学院,合肥230039 [2]安徽农业大学动物科技学院,合肥230036 [3]阜阳师范学院生命科学学院,阜阳236041

出　　处：《生物学杂志》2010年第3期5-7,共3页Journal of Biology

基　　金：国家自然科学基金资助(编号:30570922 30871415)

摘　　要：实验通过对兔子基因组DNA进行PCR获得兔IFRG基因,将其克隆到pGEM-T载体,双酶切鉴定和测序结果表明成功构建了重组克隆载体pGEM-T-IFRG。通过RT-PCR分析发现IFRG基因在兔不同组织中有不同程度的表达。将重组克隆载体pGEM-T-IFRG和表达载体pET-41(c)经过EcoR1和Xho1双酶切后连接构建重组表达载体pET-41(c)-IFRG,将双酶切鉴定正确的重组表达载体转化入E.coli BL21,IPTG诱导融合蛋白的表达,SDS-PAGE结果显示兔IFRG基因在大肠杆菌中得到了良好的表达。The rabbit IFRG gene was gained by PCR from the rabbit genomic DNA and the gene was cloned into pGEM-T vector.The double enzymes digestion and sequence analysis showed that the construction of recombinant vector pGEM-T-IFRG was successful.RT-PCR analysis of rabbit different organism showed that IFRG gene expressed distinct in different organism.To construct recombinant expression vector pET-41（c）-IFRG by the double enzymes digestion of recombinant pGEM-T-IFRG and pET-41（c）,the correct recombinant expression vector that identified with double enzymes digestion was transformed into E.coli BL21（DE3）.Fusional protein was expressed under IPTG induction.SDS-PAGE showed that the rabbit IFRG gene had a favourable expression in bacterium.

关 键 词：IFRG基因 基因克隆 RT-PCR 原核表达 

分 类 号：Q785[生物学—分子生物学] Q786