A total of 288 grapevine samples of 61 different grapevine cultivars,collected from 22 provinces and regions,were analyzed by reverse transcription-PCR(RT-PCR) for the presence of grapevine fabavirus(GFabV).PCR detect...A total of 288 grapevine samples of 61 different grapevine cultivars,collected from 22 provinces and regions,were analyzed by reverse transcription-PCR(RT-PCR) for the presence of grapevine fabavirus(GFabV).PCR detection results showed the incidences of GFabV were 12.8%(30/235) and 48.1%(25/52) in the asymptomatic and symptomatic vines,respectively.The genetic diversity of GFabV isolates was analyzed based on partial nucleotide and encoded amino acid sequences of the RNA1 and RNA2 polyprotein genes.Phylogenetic analyses of the RNA1 and RNA2 gene sequences divided the GFabV isolates into five well-defined groups.Groups 1,2,and 4 comprised only Chinese isolates.This article represents the first report for the prevalence and genetic diversity of GFabV in grapevines grown in China.展开更多
Very virulent infectious bursal disease virus(vvIBDV)was isolated from chicken bursa and then the virus double stranded RNA was extracted After denaturation of dsRNA by heat in the presence of primers,the cDNA was syn...Very virulent infectious bursal disease virus(vvIBDV)was isolated from chicken bursa and then the virus double stranded RNA was extracted After denaturation of dsRNA by heat in the presence of primers,the cDNA was synthesized by use of a reverse transcriptase lacking RNase H activity The RNA component of RNA cDNA hybrids was digested by RNase H By using an optimized PCR,a 3 05kb DNA fragment coding for precursor polyprotein of IBDV was produced in one step,which was inserted into pcDNA3 1(+) vector Two recombinant plasmids (pPP1 and pPP2)were screened and identified from eight XL1 blue colonies Partial sequencing for pPP1 plasmid indicated that the precursor polyprotein gene for IBDV was cloned successfully The method can simplify greatly the procedure to clone precursor polyprotein gene of IBDV展开更多
基金supported by the National Key R&D Program of China(2018YFD0201301)the earmarked fund for the China Agriculture Research System(CARS-29-bc-1)the Fundamental Research Funds for Central Non-profit Scientific Institutions,China
文摘A total of 288 grapevine samples of 61 different grapevine cultivars,collected from 22 provinces and regions,were analyzed by reverse transcription-PCR(RT-PCR) for the presence of grapevine fabavirus(GFabV).PCR detection results showed the incidences of GFabV were 12.8%(30/235) and 48.1%(25/52) in the asymptomatic and symptomatic vines,respectively.The genetic diversity of GFabV isolates was analyzed based on partial nucleotide and encoded amino acid sequences of the RNA1 and RNA2 polyprotein genes.Phylogenetic analyses of the RNA1 and RNA2 gene sequences divided the GFabV isolates into five well-defined groups.Groups 1,2,and 4 comprised only Chinese isolates.This article represents the first report for the prevalence and genetic diversity of GFabV in grapevines grown in China.
文摘Very virulent infectious bursal disease virus(vvIBDV)was isolated from chicken bursa and then the virus double stranded RNA was extracted After denaturation of dsRNA by heat in the presence of primers,the cDNA was synthesized by use of a reverse transcriptase lacking RNase H activity The RNA component of RNA cDNA hybrids was digested by RNase H By using an optimized PCR,a 3 05kb DNA fragment coding for precursor polyprotein of IBDV was produced in one step,which was inserted into pcDNA3 1(+) vector Two recombinant plasmids (pPP1 and pPP2)were screened and identified from eight XL1 blue colonies Partial sequencing for pPP1 plasmid indicated that the precursor polyprotein gene for IBDV was cloned successfully The method can simplify greatly the procedure to clone precursor polyprotein gene of IBDV
