sTRAIL基因的克隆及其在大肠杆菌中的表达

Clone of Gene sTRAIL and Induced Expression in E.coli

克隆TRAIL基因的 95 - 2 81位 (sTRAIL) ,构建表达载体 ,建立原核表达体系 ,优化诱导表达的条件。分离人外周血淋巴细胞 ,提取总RNA ,RT PCR ,克隆sTRAIL的cDNA ,构建原核表达载体 ,优化IPTG诱导的条件。结果 :(1)克隆了TRAIL基因 95 2 81位的cDNA ,DNA测序结果与报道的一致。 (2 )构建了sTRAIL基因的原核表达载体 ,酶切结果与预期的一致。 (3)转化宿主菌 ,优化IPTG诱导条件 ,发现 3mmol L ,诱导 3～ 4h表达最好。sTRAIL基因的克隆及表达为下游中试发酵及纯化奠定了上游的基础 ,也为研究TRAIL抗肿瘤的机制提供了可能。

amino acid of TRAIL(sTRAIL) was cloned ,constructed expressing vector. We Separated human peripheral blood, abstracted total RNA, RT-PCR ,cloned sTRAIL cDNA ,constructed prokaryotic expression vector and optimized the condition of induction. Result(1) 95-281 amino acid of TRAIL cDNA was cloned, and the DNA sequence was coincide with reported document.(2) The prokaryotic expression vector was constructed and enzyme cut confirmed the vector was constructed successful. (3) Transferred host and optimized the condition of induction. The condition that 3mmol/L IPTG and 3-4h induction are considered as the best. sTRAIL gene clone and expression provide the base of upstream for middle-scale product and for researching the mechanism of TRAIL resist tumor.