rhIL-18在不同原核表达系统中的表达和活性比较  

The Expression and Activity of rhIL-18 in Different Proaryote Express System

作　　者：张海峰[1] 李丽娥[1] 盛伟华[1] 缪竞诚[1] 谢宇锋[1] 王金志[1] 杨吉成[1] 

机构地区：[1]苏州大学医学院细胞和分子生物学教研室,江苏苏州215007

出　　处：《苏州大学学报（医学版）》2006年第1期4-6,共3页Suzhou University Journal of Medical Science

基　　金：江苏省高校自然科学研究项目(01KJB180001)

摘　　要：目的研究rhIL-18包涵体、可溶性(His融合型)两种形式蛋白的生物学活性。方法从已构建的pBV220-IL-18重组质粒中双酶切获取IL-18基因,回收后重组至pLHis质粒构建重组载体。42℃热诱导pBV220-IL-18表达,产生rhIL-18包涵体,IPTG诱导pLHis-IL-18表达,产生可溶性rhIL-18蛋白。两者分别刺激人外周血单个核细胞,ELISA法检测上清中IFN-γ的含量。结果两种重组表达载体均表达产生了rhIL-18,且均可刺激人PMBC上清产生IFN-γ,可溶性rhIL-18刺激人外周血产生IFN-γ的能力高于包涵体形式的rhIL-18。结论可溶性rhIL-18具有比包涵体形式的rhIL-18更强的活性,这为IL-18原核表达基因工程药物的开发提供了新思路。Objective To study the biologic activity of recombinant human IL-18, including inclusion body and soluble protein（His fused）. Methods The cDNA of IL-18 was obtained from recombinant plasmid pBV220-IL-18 by digesting with EcoRI and BamHI. The pLHis-IL-18 was constructed by cloning the eDNA of IL-18 into pLHis. The rhIL-18 inclusion body expressed and produeted by pBV220-IL-18 and rhIL-18 soluble protein by pLHis-IL-18 were introdueted by heat and IPTG respectively . Then the human periblood monocular eells（PBMCs） were stimulated by the two different form IL-18 production. And the IFN-γ content of the PBMCs supernatant was tested by ELISA. Results The E coli DHSa carrying pBV220-IL-18 or pLHis IL-18 could both synthesize rhIL-18. And the rhIL-18 could induce IFN-γ production from PMBCs. The IFN-γ level in the supernatant of PMBC stimulated by the soluble rhlL-18 was higher than that by inclusion body rhIL-18. Conelution The activity of soluble rhIL-18 is higher than that by inclusion body rhIL-18, so that gives us some new suggestion for exploitation of IL-18 gene engineering medicament.

关 键 词：IL-18 包涵体 可溶性蛋白 IFN-Γ 

分 类 号：R392.3[医药卫生—免疫学]