人淋巴细胞抑制素基因克隆与表达的研究

Construction and expression of the suppression gene of human lymphocyte

目的 获得人SPN基因,构建SPN的原核表达载体。方法 采用RT-PCR的方法从激活的人外周血淋巴细胞的总cDNA中得到1493bp的人SPN基因,NdeI、BamHI双酶切后定向克隆到原核表达载体pET-11-c中,全自动测序仪测序,转化宿主菌BL21后,IPTG诱导,SDS-PAGE电泳分析。结果 成功克隆到人SPN基因,并构建表达质粒,SDS-PAGE电泳证实目的蛋白表达。结论 为进一步研究SPN的免疫抑制机理和作用以及探讨SPN作为新型生理性免疫抑制剂的可能性打下了坚实的基础。

ve To obtain and construct Prokaryotic expression vector of human lymphocyte suppression gene. Methods A 1493 bp cDNA fragment was amplified by RT-PCR method from total RNA of human peripheral blood lymphocyte cell activated with 160 U/mL for 12 hours. The fragment cloning into pET-11-c plasmids.The cloned insert was identified by double I digestion of the recombinant plasmid with restriction enzymes Nde T and BamH I and sequenced by Sangers-dideory-mediated chain termination. Results This cDNA fragment in-cluded 1 493 bp entire coding region. The recombinant Prokaryotic expression vector of SPN gene was constructed and the protein of SPN was expressed . Conclusion The recombinant Prokaryotic expression vector was successfully constructed. These results pave the way for researches of SPN.