J4

• 基础研究 • 上一篇    下一篇

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

宋忆淑1,宋志宇2,费向东,贺 冰3,李玉新1*,谭大鹏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:2004-07-28 修回日期:1900-01-01 出版日期:2004-11-28 发布日期:2004-11-28
  • 通讯作者: 李玉新

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

SONG Yi-shu1, SONG Zhi-yu2, FEI Xiang-dong, HE Bing3, LI Yu-xin1*,TAN Da-peng1   

  1. 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:2004-07-28 Revised:1900-01-01 Online:2004-11-28 Published:2004-11-28
  • Contact: LI Yu-xin

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 cDNA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。

关键词: s-lap基因, 克隆, 分子, 逆转录聚合酶链反应, 基因表达, 质粒,

Abstract: 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.

Key words: s-lap gene, cloning, molecular, reverse transcriptasepolymerase chain reaction, gene expression, plasmids, swine

中图分类号: 

  • Q78