摘要
目的：分离编码人ｈＩＬ－１５的ｃＤＮＡ，并在大肠杆菌中克隆与表达。方法：采用ＰＨＡ刺激人外周血白细胞提取ｍＲＮＡ，并利用ＲＴ－ＰＣＲ方法获得了ｈＩＬ－１５的ｃＤＮＡ，将其定向插入ｐＵＣ１９载体中，ＤＮＡ序列分析表明分离的ｈＩＬ－１５的序列与文献报道一致。以ｐＢＶ２２０为表达载体构建并筛选出重组子ｐＢＶｈＩＬ－１５；重组子转化Ｅ．ｃｏｌｉＤＨ５α菌株，经４２℃诱导表达。结果：相对分子质量为１４０００的ｈＩＬ－１５表达量占菌体总蛋白的３０％，表达产物主要以包涵体形式存在。结论：本研究获得了序列正确的ｈＩＬ－１５ｃＤＮＡ克隆，并实现在Ｅ．ｃｏｌｉ中的高效表达。
Objectives: To isolate a cDNA clone encoding human interleukin 15 (hIL15) and express it in E.coli. Methods: hIL15 cDNA Was amplified by RTPCR from mRNA extracted from PHAactivated peripheral blood white cells. The cDNA encoding hIL15 was inserted into the plasmid pUC19 and sequenced by dideoxy termination method. The obtained hIL15 cDNA was recombined with plasmid pBV220. The recombinant plasmid was then introduced into the competent cells of E.coli DH5α by transformation and the expression was achieved. Results: The expressed hIL15, mainly recovered as inclusion bodies, amounts to 30% of total bacterial protein. Conclusion: A confirmed hIL15 clone was obtained and highly expressed in E.coli.
出处
《军事医学科学院院刊》
CSCD
北大核心
1998年第1期20-22,共3页
Bulletin of the Academy of Military Medical Sciences
