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

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。

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