鸡白细胞介素18基因的克隆与原核表达  

The Cloning and Prokaryotic Expression of Chicken Interleukin-18 Gene

作　　者：宋佰芬[1] 李晓婷[1] 崔玉东[1] 

机构地区：[1]黑龙江八一农垦大学,黑龙江大庆163319

出　　处：《安徽农学通报》2016年第17期19-21,共3页Anhui Agricultural Science Bulletin

基　　金：黑龙江农垦总局项目:鸡干扰素-γ和白细胞介素-18融合表达及应用效果初步分析(HNK125A-11-05)

摘　　要：白介素18(IL-18)最早被称为干扰素诱导因子(interferon-γ-inducing factor,IGIF),是近年发现的一种重要的免疫调节因子。该实验根据Gen Bank中已公布的鸡IL-18的基因序列,设计并合成了一对特异性引物,应用RT-PCR方法扩增出IL-18基因,并将该基因克隆到p MD18-T载体中,经过质粒酶切、PCR和测序鉴定,阳性克隆命名为Chp MD18-T-IL-18。将Chp MD18-T-IL-18双酶切后与p ET32a表达载体连接,转化到大肠杆菌BL21感受态细胞中,经IPTG诱导表达鸡IL-18蛋白,结果表明,在28k D左右出现了目的条带,经west-ern blot检测确定该蛋白获得高效表达,为进一步研究该蛋白的功能奠定了基础。Interleukin 18 （IL-18）is called interferon gamma inducing factor （IGIF）.It is a kind of important immune adjustment factor.In this paper, a pair of specific primer was designed and synthesised according to IL- 18 DNA se- quence in GenBank, IL-18 gene was amplified by RT-PCR method and was inserted into pMD 18-T vector.Positive plasmids were identified by PCR and restriction enzyme digestion methods.Then IL- 18 gene was inserted into pET32a vector.Positive plasmids were identified by PCR and restriction enzyme digestion methods and were trans- formed into escherichia coli competent cells.Recombinant E.coli were induced by IPTG and IL-18 protein was de- tected by SDS-PAGE and Western blot methods.The results showed IL- 18 protein was expressed.This would estab- lish a basis for studying the function of this protein.

关 键 词：鸡 白细胞介素18 原核表达 

分 类 号：Q786[生物学—分子生物学]