A novel real-time RT-PCR method,BMPs based real-time RT-PCR which integrated with magne-tic separation technique of bacterial magnetic particles(BMPs),was set up for detection of Squash mosaic virus(SqMV).After SqMV p...A novel real-time RT-PCR method,BMPs based real-time RT-PCR which integrated with magne-tic separation technique of bacterial magnetic particles(BMPs),was set up for detection of Squash mosaic virus(SqMV).After SqMV particles in crude sap were concentrated by BMPs,viral RNAs were released and detected by real time RT-PCR.The results indicated that BMPs based real-time RT-PCR was efficient,and the detection sensibility was equivalent to that of the Trizol based real-time RT-PCR,of which Trizol reagent was used for viral RNAs extration.Comparing to Trizol-based method,the BMPs-based method had advantages of simplicity on operation,time saving for RNA extraction and without using noxious organic chemicals.展开更多
The nucleotide sequence of coat protein(cp)gene of Sowbane mosaic virus(SoMV)was determined.The cp gene of SoMV consists of 726 nucleotides and encodes a putative protein of 241 amino acid re-sidues.Sequence compariso...The nucleotide sequence of coat protein(cp)gene of Sowbane mosaic virus(SoMV)was determined.The cp gene of SoMV consists of 726 nucleotides and encodes a putative protein of 241 amino acid re-sidues.Sequence comparison showed that SoMV was most closely related to Rubus chlorotic mottle virus compared to other sobemoviruses.A primer pair was designed for the detection of SoMV based upon the determined cp nucleotide sequence.An expected fragment with 510 bp could be obtained from SoMV,while no specific band was observed from the healthy control.The result showed that the RT-PCR assay with the primer pair was suitable for the specific detection of SoMV.展开更多
文摘A novel real-time RT-PCR method,BMPs based real-time RT-PCR which integrated with magne-tic separation technique of bacterial magnetic particles(BMPs),was set up for detection of Squash mosaic virus(SqMV).After SqMV particles in crude sap were concentrated by BMPs,viral RNAs were released and detected by real time RT-PCR.The results indicated that BMPs based real-time RT-PCR was efficient,and the detection sensibility was equivalent to that of the Trizol based real-time RT-PCR,of which Trizol reagent was used for viral RNAs extration.Comparing to Trizol-based method,the BMPs-based method had advantages of simplicity on operation,time saving for RNA extraction and without using noxious organic chemicals.
文摘The nucleotide sequence of coat protein(cp)gene of Sowbane mosaic virus(SoMV)was determined.The cp gene of SoMV consists of 726 nucleotides and encodes a putative protein of 241 amino acid re-sidues.Sequence comparison showed that SoMV was most closely related to Rubus chlorotic mottle virus compared to other sobemoviruses.A primer pair was designed for the detection of SoMV based upon the determined cp nucleotide sequence.An expected fragment with 510 bp could be obtained from SoMV,while no specific band was observed from the healthy control.The result showed that the RT-PCR assay with the primer pair was suitable for the specific detection of SoMV.
