重组人白细胞介素18基因的克隆及序列测定

从正常人外周血液中提取RNA,然后反转录cDNA,应用逆转录多聚酶链式反应（RT-PCR）技术,自行设计并合成一对特异性引物,成功地扩增出hIL-18cDNA基因,并定向插入PGEM-T载体中,对其进行酶切分析及序列测定.酶切分析表明一约481 bp的基因片断克隆至PGEM-T载体中,与理论设计的一致.测序结果表明克隆至PGEM-T中的IL-18cDNA包含成熟IL-18的全部编码序列,本研究结果将为今后hIL-18基因的表达及进一步在基因水平上探讨hIL-18的生物学功能提供物质基础.

Cloning and Sequence Analysis of Recombinant Human Interleukin-18 Gene

With reference to the human interleukin 18 gene in GeneBank database, a pair of DNA primers was designed and synthesized. A 481bp fragment containing hlL-18 gene was successfully amplified by using RT-PCR and then cloned into PGEM-T plasmid, enzyme digested by EcoR I and Sal I indicated that there is one nucleotide different from the published hlL-18 cDNA sequence. The sequence analysis indicated that in the sequence of IL-18- PGEM-T contains all the coding sequence for mature protein of hlL-18. The result would promote the further research in molecular biology of hlL-18, such as the exoression and the biological function research.