【摘要】 以猪瘟病毒(Hog Cholera Vivus,HCV)石门系毒株常规感染PK15细胞,提取感染细胞总RNA,同时根据猪瘟病毒Alfort株的基因组序列化学合成了8条引物。应用合成的3’端引物及提取的总RNA进行反转录合成第一链cDNA,再以此单链cDNA为模板,应用PCR法扩增猪瘟病毒cDNA片段,结果证明所扩增片段的长度与根据Alfort株基因组序列推测的长度相符,依次为693bp、629bp、346bp、285bp、120bp,最后按标准方法将346bp和120bp两个片段克隆到pUC19的BamHI位点。更多还原

【Abstract】 According to the nucleotide sequence of genome of HCV Alfort strain 8 primers were designed andchemically synthesized, and by reverse transcription- polymerase chain reaction (RT-PCR) , 5 cDNAfragments of the Chinese shimen HCV strain were amplified from total RNA extracted from HCV infectedPK 15 cells. Agarose gelelectrophoresis of RT-PCR products indicated that their sizes correspond to theexpected 693bp, 629bp, 346bp, 285bp and 120bp. Endonuclease digestion assay of the amplified fragmentPI/PA4 confirmed the results. Two of them with the sizes of 346bp and 120bp were cloned into pUC19vectors.更多还原