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The Construction of prk Gene Non-fusion Expression Vector and the Expression of Prk Protein in E. coli
Author(s): 
Pages: 341-343
Year: Issue:  4
Journal: ACTA UNIVERSITATIS MEDICINALIS NANJING

Keyword:  prk基因激酶基因表达非融合蛋白;
Abstract: 目的:构建prk基因的非融合表达载体,并诱导表达.方法:PCR扩增并回收prk基因片段,将其与亚克隆载体pMDl8-T重组,通过蓝/白斑筛选、酶谱分析和序列测定鉴定出重组质粒,然后从该重组质粒上切下prk基因片段,再克隆至非融合表达载体pBV220中,酶谱分析鉴定出正向重组质粒,诱导含有正向重组质粒的宿主菌表达蛋白,提取全菌总蛋白进行聚丙烯酰胺凝胶电泳(SDS-PAGE)分析.结果:成功构建并筛选出pBV220正向重组质粒,经热诱导表达,得到一可溶性的相对分子质量为65 ku的重组蛋白.结论:pBV220-prk表达载体的成功构建和表达,说明扩增所得的基因具有正确的阅读框架,使获得天然Prk蛋白成为可能,为进一步研究prk基因的生物学功能奠定了基础.
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