【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。更多还原

【Abstract】 Objective To construct clone of ⁸⁸Arg IL-2 and express it in Escherichia coli. Methods Site-specific mutated 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.更多还原