东亚三角涡虫肌球蛋白轻链融合蛋白原核表达载体的构建及其在大肠杆菌中的表达
首发时间:2010-11-15
摘要:目的:为了表达肌球蛋白轻链融合蛋白,构建了原核表达载体。方法:根据涡虫基因文库中肌球蛋白轻链(Mlc)基因完整的ORF序列设计合成特异性引物, 通过PCR扩增涡虫Mlc基因,并插入到融合蛋白原核表达载体PET-28中 ,转化宿主菌BL21(DE3)细胞,用IPTG 诱导表达MLC蛋白。结果:重组质粒测序和酶切结果显示Mlc 基因已正确插入PET-28中 ,重组蛋白SDS-PAGE 在18.2KD 处有一条明显的蛋白表达条带。western blot检测得到同样大小的条带。结论:涡虫His-MLC融合蛋白已被成功表达。
关键词: 东亚三角涡虫 肌球蛋白轻链 原核表达 western blot
For information in English, please click here
Construct Prokaryotic Expression Vector of Planarian myosin light chain fusion protein and Expression in Escherichia coli
Abstract:Objective: To construct planarian myosin light chain fusion protein, and for prokaryotic expression. Methods: According to the planarian myosin light chain (Mlc) mRNA sequence of primers were designed from the planarian genebank, PCR amplification of planarian Mlc gene, and insert prokaryotic expression vector PET-28, transformed into host strain BL21 (DE3) cells, isopropyl matte base galactose (IPTG), expression of MLC protein. Results: The recombinant plasmid DNA sequencing and restriction enzyme digestion results show that Mlc gene had been correctly inserted PET-28, the recombinant protein and SDS-PAGE in the purified product 18KD Department has a distinct protein band. Western blots get the same result . Conclusion: His-MLC fusion protein of planarian have been successfully expressed.
Keywords: Planarian Dugesia japonica myosin light chain Prokaryotic expression western
论文图表:
引用
No.4390923544970128****
同行评议
共计0人参与
勘误表
东亚三角涡虫肌球蛋白轻链融合蛋白原核表达载体的构建及其在大肠杆菌中的表达
评论
全部评论0/1000