pcDNA3.1/RPs15载体构建及对皮肤成纤维细胞作用的实验研究  

The effect of pcDNA 3.1/RPs15 on skin fibroblasts in vitro

作　　者：唐胜建[1] 王小柯[2] 吕晓杰[1] 刘霞[1] 张鹏[1] 张军[1] 周晓燕[1] 

机构地区：[1]潍坊医学院整形外科研究所,潍坊261042 [2]潍坊医学院分子生物学教研室

出　　处：《中华整形外科杂志》2005年第1期58-61,共4页Chinese Journal of Plastic Surgery

基　　金：山东省自然科学基金重点资助项目 (2 0 0 0C0 3 )

摘　　要：目的　构建核糖体蛋白s15 (RPs15 )基因真核表达载体 ,研究其对小鼠皮肤成纤维细胞的体外作用。方法　将RT PCR法获得的鼠胚皮肤RPs15cDNA克隆入真核表达载体pcDNA3 1(- ) ,在FuGENE6介导下转染体外培养的小鼠皮肤成纤维细胞 ,观察RPs15基因的表达情况及其对成纤维细胞生物学特性的影响。结果　构建的重组表达载体pcDNA3 1 RPs15经DNA测序证实序列正确。RT PCR法检测RPs15基因在转染的成纤维细胞中获得表达 ,细胞生长密度下降 ,形态发生变化。结论　成功构建了真核表达载体pcDNA3 1 RPs15 ,并可在体外培养的小鼠皮肤成纤维细胞中表达 ,为深入研究RPs15基因对皮肤成纤维细胞的作用奠定了基础。Objective To construct eukaryotic expression vector of ribosomal protein s15(RPs15)gene and study its effect on mouse skin fibroblasts in vitro. Methods The RPs15 cDNA encoding region of fetal mouse skin was amplified by reverse transcription polymerase chain reaction (RT PCR) method and cloned into eukaryotic expression vector pcDNA3.1 (-). The recombinant plasmid was transfected into adult mouse skin fibroblasts by FuGENE6 transfection reagents. Then the expression of RPs15 gene was detected and its biological effect on fibroblasts was measured. Results The DNA sequencing result of pcDNA3.1/RPs15 was identical with the reported. The RPs15 gene was expressed in transfected fibroblasts. The growth density of fibroblasts decreased with the conformation changing accordingly. Conclusions The eukaryotic expression vector pcDNA3.1/RPs15 is successively constructed and can be expressed in mouse skin fibroblasts. The results set up a basis for further study of the effect of RPs15 gene on skin fibroblasts. ;

关 键 词：成纤维细胞 实验研究 核糖体蛋白s15基因 真核表达载体 基因表达 

分 类 号：R346[医药卫生—基础医学]