人白细胞介素23 p19亚基基因的克隆和序列分析

Molecular cloning and sequence analysis genes of human interleukin-23 p19 subunit

摘要: 目的:克隆人白细胞介素-23(IL-23)p19亚基基因。方法:分离健康成年人外周血单个核细胞,常规培养后用脂多糖至终浓度为100 ng/m l刺激增殖12 h,收集细胞并提取细胞总RNA,用RT-PCR方法扩增人IL-23 p19 cDNA编码基因,克隆至pMD18-T载体,PCR鉴定后进行序列测定。结果:RT-PCR产物电泳结果显示所扩增的基因为734 bp,基因测序显示其序列与GenBank报道的人IL-23 p19基因序列完全一致。结论:成功地克隆了人IL-23 p19 cDNA编码基因。

Abstract: Objective:To clone genes for encoding human interleukin-23 (IL-23) p19 subunit.Methods:Peripheral blood mononuclear cells (PBMCs) of human were isolated and cultured for 10 hours,and then stimulated with 100 ng/ml LPS for 12 hours.Total RNA from these stimulated PBMCs were extracted,and cDNA of genes for human IL-23 p19 subunit were ampliphied by RT-PCR assay and inserted into pMD18-T vector.Results:The PCR production of human IL-23 p19 was verified by Electrophoresis proved to be 734 bp,result of sequencing was the same as the gene sequence of GenBank.Conclusions:cDNA encoding human interleukin-23 p19 was successfully cloned into cloning vector pMD18-T.