• 昆明醫(yī)學(xué)院第二附屬醫(yī)院肝膽外科（昆明650101）;

目的對(duì)單純皰疹病毒Ⅱ型胸苷激酶基因及血管生成抑制素進(jìn)行基因擴(kuò)增，克隆構(gòu)建真核表達(dá)質(zhì)粒pcDNA3/HSVⅡ TK/As。方法從單純皰疹病毒Ⅱ型（HSVⅡ）Sav株感染的Hep2細(xì)胞上清液中提取HSV2基因組DNA，以此為模板，用PCR方法擴(kuò)增胸苷激酶（TK）基因，將擴(kuò)增的片段克隆入載體pcDNA3中，挑取陽(yáng)性克隆測(cè)序。限制性核酸內(nèi)切酶酶切pcDNA3/HSVⅡTK,得到目的基因TK，將其克隆于已構(gòu)建的真核表達(dá)載體pcDNA3/As上。結(jié)果HSVⅡTK基因全部編碼區(qū)為1 128 bp，基因序列與genebank中報(bào)道相符。新質(zhì)粒pcDNA3/HSVⅡTK/As經(jīng)限制性核酸內(nèi)切酶(BamH Ⅰ,Hind Ⅲ)酶切后得到700 bp(As)及1000 bp(HSVⅡTK)相應(yīng)基因片段。結(jié)論本研究成功構(gòu)建pcDNA3/HSVⅡTK/As真核表達(dá)質(zhì)粒。

引用本文： 王琳,郭永章,李立,張小文,李曉,崔江云. 單純皰疹病毒Ⅱ型胸苷激酶基因及血管生成抑制素融合基因真核表達(dá)質(zhì)粒的構(gòu)建及鑒定. 中國(guó)普外基礎(chǔ)與臨床雜志, 2002, 9(4): 246-248. doi: 復(fù)制

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