人RB1及其突变体的表达与定位  

The localization and expression of human RB1 and its mutants

作　　者：潘林鑫[1] 李新颖[1] 徐南[1] 李克娟[1] 乔正[1] 耿慧武[1] 刘晓颖[1] 范礼斌[1] 

机构地区：[1]安徽医科大学生命科学院生物学教研室,合肥230032

出　　处：《安徽医科大学学报》2013年第8期853-858,共6页Acta Universitatis Medicinalis Anhui

基　　金：安徽省教育厅自然科学重点科研项目(编号:KJ2010A187);国家自然科学基金青年基金(编号:81201368);安徽省自然科学基金(编号:11040606M170;11040606M164);安徽医科大学博士科研基金(编号:XJ201009)

摘　　要：目的研究人RB1及其缺失突变体蛋白在细胞内的表达与定位。方法以含人RB1全长cDNA序列的质粒为模板,通过PCR方法分别扩增出RB1全长及其三段缺失突变体序列,然后分别构建其表达载体,利用Western blot法和免疫荧光法检测其在细胞株中的表达与定位。结果成功构建了pCDNA3.1-FLAG-RB1及其缺失突变体的表达载体,Western blot法结果证明其在细胞中都能有效地表达,免疫荧光的结果显示了RB1主要分布在细胞核,但其缺失突变体在细胞质中也有分布。结论人RB1在HEK 293T、COS7细胞中均有表达,且主要分布在细胞核内,其缺失突变体在胞质中也有分布,这为进一步了解人RB1的功能提供了一定的基础。Objective To investigate the expression and localization of the product of RB1 gene and its deletion mutants in cells. Methods The full length cDNA fragment of RB1 from plasmid was used to amplify with PCR, to get the sequences of wild-type RB1 and its deletion mutants , then to construct expression vectors. Immunofluores-cence and Western blot were adopted to detect the location and expression of wild-type RB1 and its deletion mutants. Results The expression vectors of pCDNA3. 1-FLAG-RBI and its deletion mutants were constructed successfully, Western blot results showed that it could be effectively expressed in the cells, and the location of RBI protein was in the nucleus, but the deletion mutants also had distribution in the cytoplasm. Conclusion The pRB1 can express ef- ficiently in HEK 293T and COS7 cell lines, and mainly localized in the nucleus, but its deletion mutants are also dis- tributed in the cytoplasm. The results provide some basis to understand the function of the human RB1.

关 键 词：RB1 转染 基因表达 细胞定位 

分 类 号：R341[医药卫生—基础医学] R394.2