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采用PCR技術(shù),從原核表達(dá)質(zhì)粒PUC19-VEGF165中獲得上下游含有Hind Ⅲ和BamH I酶切位點(diǎn)的目的基因VEGF165,與真核載體pEGFP-C1構(gòu)建重組質(zhì)粒pEGFP-VEGF165,觀察其在內(nèi)皮細(xì)胞中的表達(dá).結(jié)果 表明,帶有綠色熒光蛋白報(bào)告基因的真核表達(dá)質(zhì)粒pEGFP-VEGF165構(gòu)建成功,并在PEI的介導(dǎo)下成功轉(zhuǎn)染臍靜脈內(nèi)皮細(xì)胞(HUVEC),在熒光顯微鏡下可見強(qiáng)綠色熒光蛋白表達(dá),ELISA檢測(cè)證明VEGF在細(xì)胞上清中有效表達(dá),RT-PCR證明了VEGF165 mRNA水平上的有效表達(dá).此研究結(jié)果將為再狹窄的基因治療提供實(shí)驗(yàn)基礎(chǔ).

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