论文部分内容阅读
目的 :克隆小鼠内皮抑素 (m Endostatin)编码区 c DNA序列并构建含 Egr- 1启动子的 IFNγ和 m Endo-statin双基因表达载体。方法 :利用逆转录多聚酶链反应法 (RT- PCR) ,以小鼠肝细胞 m RNA为模板 ,扩增获得全长 m Endostatin,与 p MD1 8T载体连接作全自动测序 ,并利用基因重组技术构建含 Egr- 1启动子的 IFNγ和m Endostatin双基因表达质粒。结果 :经测序证实获得的 m Endostatin序列与文献报道完全一致 ,并构建了含 Egr-1启动子的 IFNγ和 m Endostatin双基因表达质粒 p Egr- IFNγ- m Endostatin。结论 :利用 RT- PCR法成功克隆了m Endostatin的 c DNA序列 ,构建了 p Egr- IFNγ- m Endostatin重组双基因表达质粒。
OBJECTIVE: To clone the c DNA sequence encoding mouse Endostatin and construct IFNγ and m Endo-statin double gene expression vectors containing Egr-1 promoter. Methods: Full-length m Endostatin was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) using mouse hepatocyte m RNA as a template. The full-length m Endostatin was ligated with p MD1 8T vector and sequenced by using the gene recombination technique IFNγ and m Endostatin double gene expression plasmids containing Egr-1 promoter. Results: The sequence of m Endostatin confirmed by sequencing was completely consistent with that reported in the literature. The Egr-IFNγ-m Endostatin gene expression plasmid containing Egr-1 promoter IFNγ and m Endostatin was constructed. Conclusion: The c DNA sequence of m Endostatin was successfully cloned by RT-PCR and the recombinant double gene expression plasmid p Egr-IFNγ-m Endostatin was constructed.