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目的 克隆小鼠慢性视网膜变性 /盘膜边缘蛋白基因 ,并构建其重组质粒载体.方法 以正常小鼠视网膜 RNA为模板 ,采用 RT- PCR方法扩增出 c DNA目的片段 ,PCR扩增 ,克隆至 p Bluescript II KS(+ )质粒 ,进行限制性内切酶 Bam H I和测序分析 ;再克隆到带有 CMV启动子的 pc DNA3质粒 Bam H I位点 ,Bam H I和 Eco R I限制性内切酶及测序证实.结果 限制性内切酶分析及测序证实 p Bluescript II KS(+ )质粒和 pc DNA 3质粒中插入的 1.2 kb片段 ,与小鼠慢性视网膜变性 /盘膜边缘蛋白基因全部编码区相吻合.结论 获得小鼠慢性视网膜变性 /盘膜边缘蛋白基因的全部编码区序列 ,并构建到带有CMV启动子的 pc DNA3载体中 ,为进一步研究打下基础.[眼科新进展 2 0 0 1;2 1(1)∶ 7- 11]“,”Objective To clone a mouse retinal degeneration slow/peripherin gene and construct a retinal degeneration slow/peripherin gene recombinant plasmid with promotor.Methods Using mouse retina RNA as a template and mouse peripherin specific oligonucleotides as primers, reverse transcription and polymerase chain reaction (RT PCR) were used to amplify and clone a retinal degeneration slow/peripherin cDNA fragment in the plasmid pBluescript II KS (+). The dideoxynucleotide chain termination method was used to determine the nucleotide sequence. The plasmid pcDNA3, which was modified by restriction enduclease BamH I and carried CMV promotor, ligased with an 1.2kb BamH I fragment from an retinal degeneration slow/peripherin cDNA clone, recombinant plasmid pcrds was identified by BamH I and EcoR I restriction analysis and sequence. Results A 1.2 kb retinal degeneration slow/peripherin cDNA containing the full length coding region of retinal degeneration slow/peripherin had been cloned and sequenced from the mouse. The results of restriction analysis, and sequence showed that the 1.2kb BamH I fragment from an retinal degeneration slow/peripherin cDNA clone was inserted into the vector pcDNA3 to construct an retinal degeneration slow/peripherin gene recombinant plasmid with CMV promotor.Conclusion Mouse retinal degeneration slow/peripherin cDNA is obtained, and recombinant plasmid with a retinal degeneration slow/peripherin gene and promotor was constructed. It is the basis on gene therapy research of mouse with slow retinal degeneration.