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