Cloning and expression of human TRAIL gene

AIM: To clone, express and identify human TRAIL gene. METHODS: Total RNA was extracted from human PBMC, and the whole length of TRAIL gene was obtained by RT PCR. The TRAIL gene was cloned into pGEM Teasy vector and sequenced. Then the gene was inserted into Eco RⅠ and Bam HⅠ site of pBV220 expression vector. After the recombinant bacteria was induced at 42℃ for 4-5 h,the expressed protein was analyzed by SDS PAGE and western blot. RESULTS: DNA sequencing result showed that TRAIL gene was exactly consistent with the sequence reported in GenBank. SDS PAGE analysis demonstrated that TRAIL protein was expressed in E.coli , and the molecular mass of it is 17 ku. The protein band amounted to 30% of total bacteria protein and could react with anti TRAIL antibody. CONCLUSION: TRAIL gene was successfully cloned and expressed.