人血管内皮抑素腺相关病毒载体包装质粒pSNAV-hEndostatin-CMV-EGFP的构建

Construction of Packaging Plasmid pSNAV-hEndostatin-CMV-EGFP of AAV Encoding Human Endostatin cDNA

目的:构建人血管内皮抑素(Endostatin)腺相关病毒(Adeno-associated virus,AAV)载体包装质粒pSNAV-hEndostatin-CMV-EGFP。方法:采用分子克隆技术,从pCD-sEndostatin质粒获得hEndostatincDNA,并将PCR扩增产物插入至AAV包装质粒pSNAV上,构建pSNAV-hEndostatin-CMV-EGFP的AAV重组质粒,经PCR、酶切及测序鉴定。结果:经PCR、酶切鉴定及基因测序证实AAV包装质粒pSNAV-hEndostatin-CMV-EGFP构建成功。结论:构建的AAV包装质粒pSNAV-hEndostatin-CMV-EGFP可作为rAAV-hEndostatin-EGFP表达载体的包装质粒。

Objective： To construct the packaging plasmid pSNAV-hEndostatin-CMV- EGFP of adeno-associated virus vector （AAV） encoding human Endostatin （hEndostatin） cDNA. Methods： The hEndostatin cDNA obtained from plasmid pCD-sEndostatin was subcloned into the packaging plasmid pSNAV of AAV by molecular clone ways. The recombinant plasmid pSNAV-hEndostatin-CMV-EGFP was identified by PCR analysis, restriction enzymes analysis and sequencing analysis.Results： The recombinant pSNAV-hEndostatin-CMV-EGFP was correctly constructed and confirmed by PCR analysis, restriction enzymes analysis and sequencing analysis. Conclusion： The constructed AAV-hEndostatin packaging plasmid pSNAV-hEndostatin-CMV-EGFP could be the packaging plasmid of rAAV- hEndostatin-EGFP.