丽蝇抗病毒肽Alloferon1基因串联体的构建和表达

Construction and expression of antiviral peptide Alloferon1 gene tandem from blow fly

目的:克隆和表达Alloferon1基因2串联体.方法:利用同尾酶法基因同向串联方案,在人工合成Alloferon1全基因的基础上,将Alloferon1基因由单体连接成2串联体,继而将获得的正确Alloferon1基因2串联体插入pGEX-4T-1表达载体诱导表达.结果:构建的Alloferon1基因2串联体经酶切和DNA测序分析,结果表明该基因串联体的序列与设计的序列完全相同.Alloferon1基因2串联体插入pGEX-4T-1载体后,重组菌经37℃诱导4h,SDS-PAGE分析结果显示,该串联体得到较高水平的表达.结论:Alloferon1基因2串联体的构建与表达均获得了成功.

AIM ： To clone and express a tandem of repeated gene recombinant plasmid pGEX-4T-1-（ Alloferonl ）2. METHODS： Whole Alloferonl gene was synthesized and inserted into pUC18 cloning vector. Double repeats of Alloferonl gene were obtained with isoschizomer construction method. Then the correct tandem of gene repeats was inserted into EcoRI and SalI sites of pGEX- 4T-1 expression vector. RESULTS： The sequencing and restriction enzyme digestion analysis showed that 2-repeat tandem of Alloferonl gene was connected correctly and cloned into pUC18 vector. After the recombinant bacteria was induced at 37℃ for 4 h, a new anticipated protein band with relative molecule mass of 32 × 10^3 was found on SDS-PAGE gel. CONCLUSION： The correct tandem of 2 repeats of Alloferonl gene is constructed and expressed in E. coli successfully.