The cDNA cloning and nucleotide sequence analysis of potato leaf roll virus intergenic region

Two specific primers were designed and synthesized according to the genomic sequence ofPLRV reported in literature,The first strand of cDNA was synthesized by reverse-transcriptionusing RNA of PLRV Chinese isolate(PLRV-Ch)as a template,followed by PCR amplification.The synthesized cDNA was cloned into plasmid pUC19 in DH5α.The cDNA clone was further i-dentified by PCR,restriction enzymes analysis and nucleotide sequence analysis.Tests show thatthe intergenic region of PLRV-Ch isolate consists of 197 nucleotides,and there is a high homologyin nucleotide sequence in comparision with PLRV Netherland isolate(PLRV-N),PLRV Canadianisolate(PLRV-C),PLRV Austrilian isolate (PLRV-A)and PLRV Scottish (PLRV-S)The rate ofnucleotide sequence homobgy is 99%,98%, 93%and 98%,respectively.