J4

• 基础研究 • 上一篇    下一篇

s-lap/GFP融合蛋白在Hela细胞中的表达与定位

宋忆淑1,宋志宇2,费向东,贺 冰3,李玉新1*   

  1. 1. 东北师范大学遗传与细胞研究所,吉林 长春130024;2.吉林大学第一医院泌尿外科,吉林 长春130021;3.吉林大学第一医院眼科,吉林 长春130021
  • 收稿日期:2004-08-27 修回日期:1900-01-01 出版日期:2005-01-28 发布日期:2005-01-28
  • 通讯作者: 李玉新

Expression and localization of s-lap/GFPfusion protein in Hela cell lines

SONG Yi-shu1, SONG Zhi-yu2, FEI Xiang-dong, HE Bing3, LI Yu-xin1*   

  1. 1.Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024,China;2.Department of Urology, First Hospital, Jilin University, Changchun 130021, China;3. Department of Ophthalmology, First Hospital, Jilin University, Changchun 130021, China
  • Received:2004-08-27 Revised:1900-01-01 Online:2005-01-28 Published:2005-01-28
  • Contact: LI Yu-xin

摘要: 目的:观察s-lap/GFP融合蛋白在Hela细胞中的表达及定位。 方法:根据s-lap基因的编码区序列, 应用PCR技术突变其3′末端终止密码子, 并通过基因重组技术将其与GFP基因融合,构建s-lap/GFP重组质粒;采用脂质体转染法, 将s-lap/GFP融合基因转入Hela细胞中, 用荧光显微镜观察其表达。 结果:s-lap/GFP重组质粒序列测定的结果与预期设计完全一致。荧光显微镜显示, 在转染GFP基因的Hela细胞中,荧光呈网状均匀分布于细胞浆内,而细胞核内低表达;在转染s-lap/GFP融合基因的Hela细胞中,荧光均匀分布于整个细胞,尤其以细胞核内高表达。 结论:s-lap/GFP融合蛋白可在人Hela细胞中表达,并主要定位于细胞核。

关键词: GFP融合蛋白, 重组融合蛋白质类, 遗传学, 基因融合, 方法, Hela细胞

Abstract: Objective To study the expression and localization of s-lap/GFP fusion protein in Hela cell lines. Methods According to the coding area sequence of s-lap gene, stop codon located at 3′terminus of s-lap gene was mutated by PCR and fused with GFP gene. Then the s-lap/GFP fusion gene was transfected into Hela cells. The expression of s-lap/GFP fusion protein was observed under fluorescence microscope. Results DNA sequencing demonstrated that s-lap/GFP recombinant vector was constructed successfully and its fusion protein was expressed in Hela cells. The observation under fluorescence microscope showed that s-lap/GFP fusion protein was spread in entire cells, but expressed highly in the nucleus. Conclusion The s-lap/GFP fusion protein can express in Hela cells and locate in entire cells, but express highly in the nucleus.

Key words: GFP fusion protein, recobinant fusion proteins, genetics, gene fusion, methods, Hela cells

中图分类号: 

  • Q78