The cDNA of BYDV-GAV read-through protein (RTP) gene was amplified from the extracted RNA of BYDV-GAV by using the polymerase chain reaction (PCR), and cloned into pGEM-7zf( + ). Its complete nucleotide sequence was d...The cDNA of BYDV-GAV read-through protein (RTP) gene was amplified from the extracted RNA of BYDV-GAV by using the polymerase chain reaction (PCR), and cloned into pGEM-7zf( + ). Its complete nucleotide sequence was determined by dideoxynucleotide chain-termination method. The BYDV-GAV RTP gene consists of 1377nt. Its sequences were most similar to that of the RTP gene of BYDV - MAV with identities of 87.4% and 87.1% at the nucleotide and amino acid levels, respectively.展开更多
The IPTG inducible expression vector containing the BYDV GAV coat protein gene was constructed and transferred into E.coli BL21(DE3).High level expression of the specific protein was achieved by IPTG induction.The res...The IPTG inducible expression vector containing the BYDV GAV coat protein gene was constructed and transferred into E.coli BL21(DE3).High level expression of the specific protein was achieved by IPTG induction.The results of SDS PAGE and Western blottong show that the expression product which accumulates 19.5% of the total cellular proteins estimated by scanning is 24 kD BYDV GAV coat protein plus eleven amino acids of pET 5a.展开更多
The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV gen...The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV genome were similar to those of BYDV PAV-aus. The nucleotide and deduced amino acid sequences of the six ORFs were aligned and compared with those of other luteoviruses. The results showed that there was a high degree of identity between BYDV-GAV and MAV-PS1 in all ORFs except ORF5 and ORF6, which had only 87.4% and 70.2% identities respectively. The reported genomic nucleotide sequence of MAV was shorter than that of BYDV-GAV, but the comparison of the genomic nucleotide sequences for MAV-PS1 and GAV showed 90.4% sequence identity for the same region of the genome. Ac-cording to the level of sequence similarities, BYDV-GAV should be closely related to BYDV-MAV.展开更多
Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion ave...Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae, two aphid species transmitting BYDV-GAV. P50 in the extracts of S. graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation. After feeding the antiserum through membrane, the transmission efficiencies of S. graminum and S. avenae for BYDV-GAV decreased significantly. It was suggested that P50 should be related with transmission pro- cess. Location of P50 was found at the plasma membrane surrounding the accessory salivary gland (ASG) in the head tissues of S. graminum by immunogold-labelling experiment. The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.展开更多
文摘The cDNA of BYDV-GAV read-through protein (RTP) gene was amplified from the extracted RNA of BYDV-GAV by using the polymerase chain reaction (PCR), and cloned into pGEM-7zf( + ). Its complete nucleotide sequence was determined by dideoxynucleotide chain-termination method. The BYDV-GAV RTP gene consists of 1377nt. Its sequences were most similar to that of the RTP gene of BYDV - MAV with identities of 87.4% and 87.1% at the nucleotide and amino acid levels, respectively.
文摘The IPTG inducible expression vector containing the BYDV GAV coat protein gene was constructed and transferred into E.coli BL21(DE3).High level expression of the specific protein was achieved by IPTG induction.The results of SDS PAGE and Western blottong show that the expression product which accumulates 19.5% of the total cellular proteins estimated by scanning is 24 kD BYDV GAV coat protein plus eleven amino acids of pET 5a.
基金This work was supported by the National Key Basic Research of China(973 contract TG2000016201)the National Natural Science Foundation of China(Grant No.39970034).
文摘The complete nucleotide sequence of genomic RNA of BYDV-GAV was determined. It comprised 5685 nucleotides and contained six open reading frames and four un-translated regions. The size and organization of BYDV-GAV genome were similar to those of BYDV PAV-aus. The nucleotide and deduced amino acid sequences of the six ORFs were aligned and compared with those of other luteoviruses. The results showed that there was a high degree of identity between BYDV-GAV and MAV-PS1 in all ORFs except ORF5 and ORF6, which had only 87.4% and 70.2% identities respectively. The reported genomic nucleotide sequence of MAV was shorter than that of BYDV-GAV, but the comparison of the genomic nucleotide sequences for MAV-PS1 and GAV showed 90.4% sequence identity for the same region of the genome. Ac-cording to the level of sequence similarities, BYDV-GAV should be closely related to BYDV-MAV.
基金This work was supported by the National Key Basic Research of China(Grant No.TG2000016201)the National Natural Science Foundation of China(Grant No.30070498).
文摘Using 2-D electrophoresis and virus overlay assay, a 50-kDa protein (P50) exhibiting specific binding to purified virus particles of BYDV-GAV was found in the protein extracts from Schizaphis graminum and Sitobion avenae, two aphid species transmitting BYDV-GAV. P50 in the extracts of S. graminum was isolated by preparation electrophoresis and electro-eluted proteins from the gel slices for antiserum preparation. After feeding the antiserum through membrane, the transmission efficiencies of S. graminum and S. avenae for BYDV-GAV decreased significantly. It was suggested that P50 should be related with transmission pro- cess. Location of P50 was found at the plasma membrane surrounding the accessory salivary gland (ASG) in the head tissues of S. graminum by immunogold-labelling experiment. The ascertainment of the protein associated with virus transmission has a significance influence on further understanding the transmission mechanism and genetic engineering for resistant to vector transmission.
