Ⅰ型单纯疱疹病毒UL12基因在大肠杆菌BL21和Rosetta菌中的差异表达
The differential expression of HSV-1 UL12 gene in E.coli BL21 and E.coli Rosetta
作者：陈恬;李学东;代富英;张磊;赖升凤;余小平;曹康;
Author：
收稿日期： 年卷（期）页码：2012,27(01):-13-16
期刊名称：华西药学杂志
Journal Name：WEST CHINA JOURNAL OF PHARMACEUTICAL SCIENCES
关键字：HSV-1碱性核酸酶;克隆;原核表达;生物学活性
Key words：
基金项目：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)
中文摘要
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。
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