^(88)Arg IL-2的克隆构建及原核表达  

CLONING AND PROKARYOTIC EXPRESSION AND ACTIVITY OF ^(88)Arg IL-2

作　　者：孟林[1] 刘明军[1] 王斌[1] 钱冬萌[1] 

机构地区：[1]青岛大学医学院微生物学教研室,山东青岛266021

出　　处：《青岛大学医学院学报》2006年第4期333-335,338,共4页Acta Academiae Medicinae Qingdao Universitatis

摘　　要：目的构建88Arg IL-2的重组克隆,并在大肠杆菌中表达。方法根据天然IL-2的基因序列设计含目的突变位点的引物,经PCR定点诱变技术获得88Arg IL-2的重组克隆,插入原核表达质粒pGEX4T-2中,经IPTG诱导在大肠杆菌DH5α中表达88Arg IL-2与GST的融合蛋白。结果所表达的蛋白相对分子质量约为42 000。Western blot检测结果表明,该蛋白具有与亲本IL-2相同的抗原性。MTT比色结果表明,88Arg IL-2可促使IL-2依赖细胞株CTLL-2生长,具有与亲本IL-2相近的生物学活性。结论成功构建了88Arg IL-2的重组克隆,在原核表达系统中高效表达出融合蛋白,并且具有生物学活性。ObJective To construct clone of ^88Arg IL-2 and express it in Escherichia coll. Methods Site specific murated primers were designed according to natural IL-2 DNA sequence. ^88Arg IL-2 gene was amplified using PCR. Then the ^88Arg IL-2 gene was inserted into expression vector pGEX4T-2 and induced by IPTG to express the protein in E. coli DH5α. Results The molecular weight of fusion protein was 42 000. Western blotting test revealed that it had good specificity to anti IL-2. The result of MTT assay indicated that ^88Arg IL-2 could stimulate the proliferation of CTLL-2 which was IL-2-dependent cell strain. Conclusion ^88Arg IL-2 has been cloned; the fusion protein is expressed in prokaryotic expression system with biologic activity.

关 键 词：白细胞介素2 基因突变 原核表达 蛋白质印迹法 MTT比色法 

分 类 号：Q78[生物学—分子生物学]