Amplification of Full-length cDNA of Papaya Ringspot Virus of Hainan Strain
To determine the complete nucleotide sequence of Papaya ringspot virus of Hainan isolate,an efficient method was developed to amplify the full-length cDNA of PRSV.Total RNA was extracted by RNAprep Pure Plant Kit from susceptible papaya leaves.The first strand synthesis using Oligo-dT and random nine primers was carried out,which catalyzed by AMV Reverse Transcriptase.The full-length cDNA of PRSV was amplified by using an optimized PCR.Both A and B segment were generated and cloned using the amplified full-length cDNA segment as the template.The sequence of the full-length of PRSV of Hainan isolate was given in this paper.