摘要
用ＲＴ－ＰＣＲ技术从健康人外周血单核细胞的总ＲＮＡ中扩增出编码白细胞介素１８的全长ｃＤＮＡ，并将此基因定向克隆入真核表达质粒载体ｐｃＤＮＡ３中，通过对转化子的筛选得到了带有ＩＬ－１８插入片段的阳性克隆，经酶切分析及核苷酸测序表明，克隆到的基因与文献报道的完全一致。把重组质粒ｐｃＤＮＡ３／ＩＬ－１８分别转染人肝癌细胞ＨｅｐＧ２和鼠肉瘤细胞Ｓ１８０，在ｍＲＮＡ水平检测到ＩＬ－１８的表达，但ＩＬ－１８ 的表达未诱导肿瘤细胞产生ＩＦＮ－γ。
The cDNA that encoding the human interleukin-18 was cloned using a RT-PCR from human blood monocytes, and an eukaryotic expression plasmid pcDNA3/hIL-18 was constructed. Restricted enzyme and sequence analysis had proved the correctness of the cDNA and the expres- sion vector. IL-18 expresses in human hepatoma cell line HepG2 and murine sarcoma cell line S180 transfectants, which did not stimulate IFN-γ production in these cells,as measured with RT- PCR.
出处
《生物技术通讯》
CAS
1999年第4期254-257,共4页
Letters in Biotechnology
基金
全军"九五"医药卫生基金资助
