A pair of oligonucleotide primers were designed to amplify the endoflagella gene of L. Interrogans serovar lai. An approximately 840 bp fragment was generated with PCR and inserted into plasmid pUC8,after the fragment...A pair of oligonucleotide primers were designed to amplify the endoflagella gene of L. Interrogans serovar lai. An approximately 840 bp fragment was generated with PCR and inserted into plasmid pUC8,after the fragment and pUC8 were digested respectively with BamHI and PstI. A recombinant plasmid (designated as pLF1 ) was obtained. SDS-PAGE analysis indicated that a 33 kDa was expressed in E. Coli JM103 harboring pLF1 and the expression level of the protein was 11 % of the total bacterial soluble proteins. Western blot analysis showed that the protein band could be recognized by the antiserum against the endoflegella (Axial filament ) of Leptospira interrt,gans serover lai. Nucleotide sequence data showed an open reading frame encoding 282 aminoacids residues,corresponding to a protein of molecular weight 33. 6kDa. Comparison of the deduced endoflagellar subunit protein (flaB) amino acid sequence with flagellins from other bacteria revealed a high level of identity with the Trehoema pallid um flaB proteins. Immunization/protection experiment was performed on the model of BALB/C mice and showed that there was higher survival rate in the group JM103-pLE1 than in the group JM103-pUC8.展开更多
The objective of this paper was to analyze the E2 genetic characterization of HB-DCZ strain of Bovine viral diaxrhca Virus （BVDV） which wcrc amplified by RT-PCR and isolated from China. The product of PCK was cloned...The objective of this paper was to analyze the E2 genetic characterization of HB-DCZ strain of Bovine viral diaxrhca Virus （BVDV） which wcrc amplified by RT-PCR and isolated from China. The product of PCK was cloned into pMD18-T vector, and then transfected Escherichia Coli JMI00. The recombinant plasmids were amplified by PCR and were sequenced. From the nucleotide sequence of the amplified products, phylogenetie analyses were performed and genotypes or subgenotypes were identified. The results indicated that the E2 gene fragment of HB-DCZ strain contained 1277bp nucleotides, and had 89.4%, 70.7%, 97.6%, 68.9%, 67.2% sequence similarity with Osloss, OregonC24V, Changchun184, ZM195, NADL, respectively. In conclusion, HB-DCZ strain is closely related to BVDV Osloss, Changchun184, and belongs to subgenotype lb.展开更多
文摘A pair of oligonucleotide primers were designed to amplify the endoflagella gene of L. Interrogans serovar lai. An approximately 840 bp fragment was generated with PCR and inserted into plasmid pUC8,after the fragment and pUC8 were digested respectively with BamHI and PstI. A recombinant plasmid (designated as pLF1 ) was obtained. SDS-PAGE analysis indicated that a 33 kDa was expressed in E. Coli JM103 harboring pLF1 and the expression level of the protein was 11 % of the total bacterial soluble proteins. Western blot analysis showed that the protein band could be recognized by the antiserum against the endoflegella (Axial filament ) of Leptospira interrt,gans serover lai. Nucleotide sequence data showed an open reading frame encoding 282 aminoacids residues,corresponding to a protein of molecular weight 33. 6kDa. Comparison of the deduced endoflagellar subunit protein (flaB) amino acid sequence with flagellins from other bacteria revealed a high level of identity with the Trehoema pallid um flaB proteins. Immunization/protection experiment was performed on the model of BALB/C mice and showed that there was higher survival rate in the group JM103-pLE1 than in the group JM103-pUC8.
文摘The objective of this paper was to analyze the E2 genetic characterization of HB-DCZ strain of Bovine viral diaxrhca Virus （BVDV） which wcrc amplified by RT-PCR and isolated from China. The product of PCK was cloned into pMD18-T vector, and then transfected Escherichia Coli JMI00. The recombinant plasmids were amplified by PCR and were sequenced. From the nucleotide sequence of the amplified products, phylogenetie analyses were performed and genotypes or subgenotypes were identified. The results indicated that the E2 gene fragment of HB-DCZ strain contained 1277bp nucleotides, and had 89.4%, 70.7%, 97.6%, 68.9%, 67.2% sequence similarity with Osloss, OregonC24V, Changchun184, ZM195, NADL, respectively. In conclusion, HB-DCZ strain is closely related to BVDV Osloss, Changchun184, and belongs to subgenotype lb.
