Partial sequence(314 bp) of ASPV was cloned and used as a probe labelled with digoxigenin-11dUTP.The total RNA extracted from samples with Apple stem pitting virus and a series of dilutions of plasmid with ASPV-cDNA w...Partial sequence(314 bp) of ASPV was cloned and used as a probe labelled with digoxigenin-11dUTP.The total RNA extracted from samples with Apple stem pitting virus and a series of dilutions of plasmid with ASPV-cDNA were detected by dot blot hybridization.The results showed that the probe was sensitive and specific.The probe couldn’t hybridize with total RNA of Apple stem grooving virus,Apple mosaic virus and Apple chlorotic leaf spot virus samples as well as negative control,only hybridized with that extracted from dormant shoot infected with ASPV.The sensitivity for detection of plasmid contained ASPV-cDNA was 1.64 μg.展开更多
Young leaves and shoot with Apple stem pitting virus（ASPV） detected by RT-PCR（reverse transcription-polymerase chain reaction） from Korla pear were used as materials and total RNA was extracted.Two expected fragme...Young leaves and shoot with Apple stem pitting virus（ASPV） detected by RT-PCR（reverse transcription-polymerase chain reaction） from Korla pear were used as materials and total RNA was extracted.Two expected fragments had been obtained by RT-PCR with two specific pairs of primers based on the sequence of NC003462 published in GenBank.The specific fragments had been recovered and cloned,and the white colonies had been screened out.After enzyme digestion of the plasmids isolated,the positive clone was selected to be sequenced.Compared with NC003462,three ORFs（open reading frame）-ORF2,ORF3 and ORF4 of ASPV with 673 bp,365 bp,276 bp representing homology of 77%,88.52%,86.33% respectively had been obtained.展开更多
文摘Partial sequence(314 bp) of ASPV was cloned and used as a probe labelled with digoxigenin-11dUTP.The total RNA extracted from samples with Apple stem pitting virus and a series of dilutions of plasmid with ASPV-cDNA were detected by dot blot hybridization.The results showed that the probe was sensitive and specific.The probe couldn’t hybridize with total RNA of Apple stem grooving virus,Apple mosaic virus and Apple chlorotic leaf spot virus samples as well as negative control,only hybridized with that extracted from dormant shoot infected with ASPV.The sensitivity for detection of plasmid contained ASPV-cDNA was 1.64 μg.
文摘Young leaves and shoot with Apple stem pitting virus（ASPV） detected by RT-PCR（reverse transcription-polymerase chain reaction） from Korla pear were used as materials and total RNA was extracted.Two expected fragments had been obtained by RT-PCR with two specific pairs of primers based on the sequence of NC003462 published in GenBank.The specific fragments had been recovered and cloned,and the white colonies had been screened out.After enzyme digestion of the plasmids isolated,the positive clone was selected to be sequenced.Compared with NC003462,three ORFs（open reading frame）-ORF2,ORF3 and ORF4 of ASPV with 673 bp,365 bp,276 bp representing homology of 77%,88.52%,86.33% respectively had been obtained.
