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新城疫病毒特异性糖蛋白基因探针的制备及应用
引用本文:刘伟忠,姜焱,吴艳涛,张如宽,刘秀梵.新城疫病毒特异性糖蛋白基因探针的制备及应用[J].畜牧兽医学报,1999,30(1):44-49.
作者姓名:刘伟忠  姜焱  吴艳涛  张如宽  刘秀梵
作者单位:扬州大学农学院动物医学系
基金项目:江苏省自然科学基金,教委自然科学基金
摘    要:应用RT-PCR获取新城疫病毒F48E8株的部分囊膜糖蛋白基因片段。扩增产物经琼脂糖凝胶电泳分析,长为926bp,与预期结果相符;将该基因片段定向克隆入质粒载体PUC19只,得到重组质粒P926,酶切分析结果与已发表的NDV毒株酶切位点一致。

关 键 词:新城疫病毒  糖蛋白  基因探针

PREPARATION AND APPLICATION OF THE SPECIFIC PROBE FOR THE GLYCOPROTEIN GENES OF NEWCASTLE DISEASE VIRUS
Liu Weizhong,Jiang Yan,Wu Yantao,Zhang Rukuan,Liu Xiufan.PREPARATION AND APPLICATION OF THE SPECIFIC PROBE FOR THE GLYCOPROTEIN GENES OF NEWCASTLE DISEASE VIRUS[J].Acta Veterinaria et Zootechnica Sinica,1999,30(1):44-49.
Authors:Liu Weizhong  Jiang Yan  Wu Yantao  Zhang Rukuan  Liu Xiufan
Abstract:A cDNA fragment of newcastle disease virus(NDV)strain F48E8 containing partial fusion(F)gene and partial haemagglutinin neuraminidase(HN)gene was obtained by RT PCR.The amplified fragment was 926 bps long as expected analyzed by agarose gel electrophoresis.It was directly cloned into plasmid vector pUC19 and the positive clone named as p926 was confirmed by restriction endonuclease analysis.Then the cDNA fragment was labeled as a DIG probe for detection of the envelope glycoprotein genes of NDV.The specificity of the DIG probe was tested by RNA dot blot hybridization.The probe detected the genomic RNA of three NDV strains(F48E8,La Sota,Ulster),and no hybridization signal was observed with RNA extracted from other avian viruses including IBDV and IBV.The clones(pF and pHN)containing envelope glycoprotein genes of NDV strain F48E8 were tested in a Southern hybridization assay with the DIG probe.The result showed that the 1.7kb long cDNA fragment of pF and the 1.93kb long cDNA fragment of pHN were positive,and further confirmed that the F and HN glycoprotein genes of NDV strain F48E8 were successively cloned.
Keywords:Newcastle  disease  virus  Glycoprotein  Gene  Probe
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