新基因s-lap编码区的克隆及其重组表达质粒PHB/s-lap的构建  

Cloning of s-lap novel gene coding area and construction of recombinant expression plasmid PHB/s-lap

作　　者：宋忆淑[1] 宋志宇[2] 费向东[3] 贺冰[4] 李玉新[1] 谭大鹏[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]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）》2004年第6期856-859,共4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 :克隆新基因 s- lap编码区序列并构建其重组表达质粒。方法 :利用逆转录聚合酶链反应 (RT-PCR )方法 ,以可见光损伤早期的培养的猪视网膜色素上皮 (RPE)细胞 ds c DNA为模板 ,扩增新基因 s- lap编码区序列 ;利用基因重组技术构建 PHB/ s- lap重组表达质粒。结果 :经测序证实获得了正确的新基因 s- lap编码区序列 ,酶切鉴定及 DNA序列分析表明已经将 s- lap基因编码区序列克隆到 PHB表达载体 ,成功地构建了 PHB/ s- lap重组表达质粒。结论 :成功地克隆了新基因 s- lap编码区序列 ,并构建了其重组表达质粒 PHB/ s- lap。Objective To clone the sequence of the cDNA of s-lap novel gene coding area and construct its expression vector. Methods The visible light damaged cultured retinal pigment epithelial (RPE) cell ds cDNA was used as template to obtain s-lap novel gene coding area sequence with RT-PCR and to construct its recombinant expression plasmid with recombinant DNA technique.Results Sequencing proved that the correct s-lap novel gene coding area sequence was obtained and s-lap gene coding area sequence was cloned to PHB expression vector by enzyme digestion identification and DNA sequence analysis,its recombinant expression vector was constructed successfully.Conclusion The s-lap novel gene coding area sequence is cloned and its recombinant expression plasmid PHB/s-lap is constructed successfully.

关 键 词：基因 s-lap基因 克隆 分子 逆转录聚合酶链反应 基因表达 质粒 猪 

分 类 号：Q78[生物学—分子生物学]