摘要
目的构建Ⅰ型单纯疱疹病毒(HSV-1)的UL12基因原核重组表达质粒pET32a-UL12,比较重组表达质粒pET32a-UL12在E.coli BL21和E.coli Rosetta表达产量与生物学活性。方法使用PCR的方法扩增出HSV-1的UL12基因,克隆至原核表达质粒pET32a(+),构建出重组质粒pET32a-UL12,分别转化E.coli BL21和E.coli Rosetta菌。经SDS-PAGE鉴定分析目的蛋白的表达差异,并比较在两种菌中目的蛋白的活性。结果重组质粒pET32a-UL12在E.coli BL21和E.coliRosetta的包涵体中均表达出了碱性核酸酶融合蛋白,E.coli Rosetta中表达的碱性核酸酶融合蛋白表达量较大,而E.coliBL21中表达的碱性核酸酶活性更高。结论成功构建了表达出了具有较高活性的HSV-1碱性核酸酶融合蛋白,为进一步研究抗HSV-1药物奠定了基础。
OBJECTIVE To construct the recombinant plasmid of pET32a-UL12 and to compare the expression level and biological activity of expression products of pET32a-UL12 in E.coli BL21 and E.coli Rosetta.METHODS The UL12 gene of HSV-1 was obtained by PCR,and was cloned into pET32a（＋）.The recombinant plasmid of pET32a-UL12 was transformed into E.coli BL21 and E.coli Rosetta.The expressed proteins of the E.coli BL21 and E.coli Rosetta were identified by SDS-PAGE.Then,the expression level and the biological activity of the two proteins were compared in this study.RESULTS The UL12 gene was successfully expressed both in E.coli BL21 and in E.coli Rosetta.The UL12 gene expressed more proteins in E.coli Rosetta than that in E.coli BL21,while the biological activity of the proteins expressed in E.coli BL21 was higher than that in E.coli Rosetta.CONCLUSION The HSV-1 alkaline nuclease with high biological activity was obtained in the study which is very important in the research of anti-virus remedies.
出处
《华西药学杂志》
CAS
CSCD
北大核心
2012年第1期13-16,共4页
West China Journal of Pharmaceutical Sciences
基金
国家自然科学基金资助项目(批准号:81173637)
四川省青年科技基金项目(06ZQ026-024)
2010年成都医学院创新性实验项目(CX2010024)
