To support the scientific basis for rapid identification of pathogenic bacteria and other stud- ies,the sequences of hsp60 gene in major 34 species of 16 genus of pathogenic bacteria were search out in GenBank and a p...To support the scientific basis for rapid identification of pathogenic bacteria and other stud- ies,the sequences of hsp60 gene in major 34 species of 16 genus of pathogenic bacteria were search out in GenBank and a proper pair of universal degenerate primer was designed by means of the molecu- lar biological softwawe Primer 5.0 and Oligo 6.0.This primer was then used in the PCR amplification, and the hsp60 gene fragments of the selected pathogenic bacteria could be amplified using this degener- ate primer.By way of bioinformational analysis,the conservation,variation and the interspecies phylo- genetic relations of the hsp60 gene sequence were analysed.From the results of the comparative study on sequences,it was demonstrated that the hsp60 gene was characterized by conservation and varia- tion,in which the conserved and mutant regions co-existed and separately distributed with many small mutant regions distributed among the conserved regions,just like the mosaic.The phylogenetic tree among different pathogenic bacteria drawn from the hsp60 gene analysis was proved to be consistent with those from 16S rRNA and 23S rRNA.It is concluded that the sequence distribution of hsp60 gene would provide a solid basis for the rapid identification of pathogenic bacteria and the development of a diagnostic microarray.展开更多
To test the antigenic activity of M protein (Mc protein) in the inner membrane of SARS-CoV, SARS-CoV Mc protein's bases locating inside the membrane were cloned, the His-fusion protein was expressed in E. coli and...To test the antigenic activity of M protein (Mc protein) in the inner membrane of SARS-CoV, SARS-CoV Mc protein's bases locating inside the membrane were cloned, the His-fusion protein was expressed in E. coli and analyzed for its antigenic activity. Among those 7 clinically diagnosed patients' sera , there were 5 positive and 2 negative in reaction with His-fusion protein. All of the 20 healthy persons' sera and rabbit anti-OC43 and 229E were of negative reaction with His-fusion protein. The animals immunized with His-fusion protein have produced multi-clonal antibody. The His-fusion protein could specially react with clinically diagnosed SARS patients' sera and the animals immunized with His-fusion protein could produce specifically multi-clonal antibody, but it could not react with the sera of healthy persons and the rabbit anti-OC43 and 229E.展开更多
文摘To support the scientific basis for rapid identification of pathogenic bacteria and other stud- ies,the sequences of hsp60 gene in major 34 species of 16 genus of pathogenic bacteria were search out in GenBank and a proper pair of universal degenerate primer was designed by means of the molecu- lar biological softwawe Primer 5.0 and Oligo 6.0.This primer was then used in the PCR amplification, and the hsp60 gene fragments of the selected pathogenic bacteria could be amplified using this degener- ate primer.By way of bioinformational analysis,the conservation,variation and the interspecies phylo- genetic relations of the hsp60 gene sequence were analysed.From the results of the comparative study on sequences,it was demonstrated that the hsp60 gene was characterized by conservation and varia- tion,in which the conserved and mutant regions co-existed and separately distributed with many small mutant regions distributed among the conserved regions,just like the mosaic.The phylogenetic tree among different pathogenic bacteria drawn from the hsp60 gene analysis was proved to be consistent with those from 16S rRNA and 23S rRNA.It is concluded that the sequence distribution of hsp60 gene would provide a solid basis for the rapid identification of pathogenic bacteria and the development of a diagnostic microarray.
文摘To test the antigenic activity of M protein (Mc protein) in the inner membrane of SARS-CoV, SARS-CoV Mc protein's bases locating inside the membrane were cloned, the His-fusion protein was expressed in E. coli and analyzed for its antigenic activity. Among those 7 clinically diagnosed patients' sera , there were 5 positive and 2 negative in reaction with His-fusion protein. All of the 20 healthy persons' sera and rabbit anti-OC43 and 229E were of negative reaction with His-fusion protein. The animals immunized with His-fusion protein have produced multi-clonal antibody. The His-fusion protein could specially react with clinically diagnosed SARS patients' sera and the animals immunized with His-fusion protein could produce specifically multi-clonal antibody, but it could not react with the sera of healthy persons and the rabbit anti-OC43 and 229E.
