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《CHINESE JOURNAL OF ANIMAL AND VETERINARY SCIENCES》 1999-01
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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 (Department of Veterinary Medicine,Agricultural College, Yangzhou University,Yangzhou,Jiangsu 225009)  
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.
【Fund】: 江苏省自然科学基金 教委自然科学基金
【CateGory Index】: S852.65
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