s-lap/GFP融合蛋白在Hela细胞中的表达与定位  

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

作　　者：宋忆淑[1] 宋志宇[2] 费向东[3] 贺冰[4] 李玉新[1] 

机构地区：[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]吉林省长春市双阳区医院骨科 [4]吉林大学第一医院眼科,吉林长春130021

出　　处：《吉林大学学报（医学版）》2005年第1期39-41,共3页Journal of Jilin University：Medicine Edition

基　　金：中国博士后科学基金资助课题 (2 0 0 4 0 35 5 5 7) ;吉林省科技厅自然科学基金资助课题 (2 0 0 30 5 4 1- 4 )

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。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.

关 键 词：s-lap/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 

分 类 号：Q78[生物学—分子生物学]