According to the published nucleotide sequences of the VP1 gene of foot-and-mouth disease virus serotype Asia 1 isolates, a pair of primers were designed and synthesized to clone the VP1 gene of YNBS/58 strain. PCR pr...According to the published nucleotide sequences of the VP1 gene of foot-and-mouth disease virus serotype Asia 1 isolates, a pair of primers were designed and synthesized to clone the VP1 gene of YNBS/58 strain. PCR product was cloned into pProexHTb vector, and E.coli BL21 was transformed by the recombinant plasmid pProex-VP1 for sequencing and expression .The expressed product was identified by SDS-PAGE and Western blot, and purified by Ni-NTA His.Bind resins. The results showed that the nucleotide sequence identity of VP1 gene between YNBS/58 and India93, India 97,India99, Iseral and YNAs1.1 strains is from 80.3% to 97.5% and amino acid identity is from 85.8% to 96.5%, the recombinant VP1 protein was significant at 34 ku by SDS-PAGE and Western blot, which accounted for 30% of total protein in E.coli lysates,and the recombinant protein was purified successfully.展开更多
The full-length genomic sequence of foot-and-mouth disease virus(FMDV) Asia1/YNBS/58 strain was determined by RT-PCR and compared with other 17 reference strains.The results showed that the complete genome of Asial/YN...The full-length genomic sequence of foot-and-mouth disease virus(FMDV) Asia1/YNBS/58 strain was determined by RT-PCR and compared with other 17 reference strains.The results showed that the complete genome of Asial/YNBS/58 was 8164nt long including a 1061-nt 5′untranslated region(UTR),a 6990-nt open reading frame(ORF),and a 113-nt 3′UTR.The homology analysis indicated that the UTR regions and non-structural proteins were more conserved than the structural proteins in FMDV.VP1 exhibited the lowest conservation and VP4 was exceptionally conserved.The VP1-,VP2-,and VP3-based phylogenetic trees were divided into distinct clusters according to different serotypes,while the other gene-based phylogenetic trees exhibited some degree of intercross among serotypes.This study is the first description of the full-length genomic sequence of FMDV Chinese serotype Asia1.展开更多
文摘According to the published nucleotide sequences of the VP1 gene of foot-and-mouth disease virus serotype Asia 1 isolates, a pair of primers were designed and synthesized to clone the VP1 gene of YNBS/58 strain. PCR product was cloned into pProexHTb vector, and E.coli BL21 was transformed by the recombinant plasmid pProex-VP1 for sequencing and expression .The expressed product was identified by SDS-PAGE and Western blot, and purified by Ni-NTA His.Bind resins. The results showed that the nucleotide sequence identity of VP1 gene between YNBS/58 and India93, India 97,India99, Iseral and YNAs1.1 strains is from 80.3% to 97.5% and amino acid identity is from 85.8% to 96.5%, the recombinant VP1 protein was significant at 34 ku by SDS-PAGE and Western blot, which accounted for 30% of total protein in E.coli lysates,and the recombinant protein was purified successfully.
文摘The full-length genomic sequence of foot-and-mouth disease virus(FMDV) Asia1/YNBS/58 strain was determined by RT-PCR and compared with other 17 reference strains.The results showed that the complete genome of Asial/YNBS/58 was 8164nt long including a 1061-nt 5′untranslated region(UTR),a 6990-nt open reading frame(ORF),and a 113-nt 3′UTR.The homology analysis indicated that the UTR regions and non-structural proteins were more conserved than the structural proteins in FMDV.VP1 exhibited the lowest conservation and VP4 was exceptionally conserved.The VP1-,VP2-,and VP3-based phylogenetic trees were divided into distinct clusters according to different serotypes,while the other gene-based phylogenetic trees exhibited some degree of intercross among serotypes.This study is the first description of the full-length genomic sequence of FMDV Chinese serotype Asia1.
