天津医药

• 论著 •    下一篇

重组真核质粒pERFP-C1-hTudor-SN-SN(1~4)的构建和表达

付晓,杨洁   

  1. 天津医科大学免疫教研室
  • 收稿日期:2010-10-12 修回日期:2010-11-19 出版日期:2011-02-15 发布日期:2011-02-15
  • 通讯作者: 付晓

Construction and Expression of pERFP-C1-hTudor-SN-SN (1~4) Recombinant Eukaryotic Plasmids

2   

  • Received:2010-10-12 Revised:2010-11-19 Published:2011-02-15 Online:2011-02-15

PDF (PC)

1954

摘要： 摘要 目的：将人类Tudor-SN蛋白SN（1~4）基因片段分别定向连入pERFP-C1质粒，使Tudor-SN蛋白SN各功能片段与红色荧光蛋白在HeLa细胞内融合表达。方法：以重组质粒pSG5-Tudor-SN-flag为模板，PCR法扩增出目的基因，利用EcoRI和BamHI双酶切法将目的片段连接到pERFP-C1载体上，再将构建成功的pERFP-C1- Tudor-SN-SN (1~4)重组质粒转染入HeLa细胞内，在荧光显微镜下观察红色荧光蛋白的表达。 结果：以单/双酶切及基因测序法鉴定构建的重组质粒均无误，荧光显微镜下观察到红色融合蛋白的表达。 结论：重组pERFP-C1- hTudor-SN-SN (1~4)质粒构建及表达成功。

关键词: 人类Tudor-SN蛋白, pERFP-C1, 重组质粒, 融合蛋白

Abstract: Abstract Objective:To construct eukaryotic red fluorescent protein(RFP) expressing recombinant plasmids, pERFP-C1-hTudor-SN-SN(1~4), which contain SN((1~4) fragments of human Tudor-SN. Methods:The genes of Tudor-SN fragments were amplified by PCR from the recombinant pSG5-Tudor-SN-flag plasmid and inserted into pERFP-C1 fluorescent expressing vector with EcoRI and BamHI site. These recombinant pERFP-C1-hTudor-SN-SN(1~4) plasmids were transfected into HeLa cells and the expression of red fluorescent fusion proteins was examined by fluorescence microscope. Results:The SN(1~4) fragments of Tudor-SN were sequenced correctly and detected in the products of the restriction single/double enzyme digestion. The red fluorescent fusion proteins were observed in HeLa cell after the transfection. Conclusion:These recombinant eukaryotic plasmids of pERFP-C1-Tudor-SN(1~4) were constructed successfully and expressed effectively. Key words Human Tudor-SN protein pERFP-C1 Recombinant plasmid Fusion protein

Key words: Human Tudor-SN protein, pERFP-C1, Recombinant plasmid, Fusion protein