Objective.To construct recombinant BCG again st leptospirosis.Methods.We amplified the entire open readin g frame of the OmpL1gene from the genome of the leptospire serovar Lai strain 017.Two recombin ant plasmids pBQ...Objective.To construct recombinant BCG again st leptospirosis.Methods.We amplified the entire open readin g frame of the OmpL1gene from the genome of the leptospire serovar Lai strain 017.Two recombin ant plasmids pBQ1and pBQ2were constructed by oriented ligation based on the E.coli-BCG shuttle plasmids pMV261and pMV361respectively.The recombinant plasmids were transformed into BCG by electroporation.The rBCGs bearing pBQ1and pBQ2were induced by high temperature of 45℃.Results.The expressed product,a 35kD prote in was detected by SDS-PAGE.The resu lt indicates that pBQ1and pBQ2can express OmpL1in rBCG.Conclusion.The technical methods in this study may help detect the immunogenicity a nd immunoprotection of OmpL1and develop more safe,highl y effective rBCG bearing leptospira l antigen with long-lasting protection.展开更多
To investigate the DNA sequence polymorphism of Chlamydia traehomatis ompl gene, urogenital samples were collected from 4 different cities in South China, DNA was extracted, and an approx- imately 980-bp-long fragment...To investigate the DNA sequence polymorphism of Chlamydia traehomatis ompl gene, urogenital samples were collected from 4 different cities in South China, DNA was extracted, and an approx- imately 980-bp-long fragment of the ompl gene was amplified by nested polymerase chain reaction （nPCR）. DNA sequence was determined, genotyping was performed by BLAST similarity search, and multiple alignment was performed with CLUSTAL X. Then a phylogenetie tree was constructed by Mega 3 software to illustrate the evolutionary relationships between clinical isolates and reference strains. Ninetysix specimens were sequenced, and 28 genetic variants were detected, among which E was the most prevalent genotype. The ompl gene was highly conserved for genotypes E and F, but appeared slightly less conserved for other genotypes, where the sequences displayed one to several nueleotide substitutions relative to the reference sequence. Phylogenetie tree showed that C. traehomatis serotypes were mainly divided into three clusters, according to previous grouping in the B, F-G, and C complexes, and the clinical isolates were highly related to the corresponding reference strains. It concluded that the ompl gene of the isolated C. traehomatis strains exhibited remarkable DNA sequence polymorphism, which can encourage for vaccine design and infection control.展开更多
文摘Objective.To construct recombinant BCG again st leptospirosis.Methods.We amplified the entire open readin g frame of the OmpL1gene from the genome of the leptospire serovar Lai strain 017.Two recombin ant plasmids pBQ1and pBQ2were constructed by oriented ligation based on the E.coli-BCG shuttle plasmids pMV261and pMV361respectively.The recombinant plasmids were transformed into BCG by electroporation.The rBCGs bearing pBQ1and pBQ2were induced by high temperature of 45℃.Results.The expressed product,a 35kD prote in was detected by SDS-PAGE.The resu lt indicates that pBQ1and pBQ2can express OmpL1in rBCG.Conclusion.The technical methods in this study may help detect the immunogenicity a nd immunoprotection of OmpL1and develop more safe,highl y effective rBCG bearing leptospira l antigen with long-lasting protection.
文摘To investigate the DNA sequence polymorphism of Chlamydia traehomatis ompl gene, urogenital samples were collected from 4 different cities in South China, DNA was extracted, and an approx- imately 980-bp-long fragment of the ompl gene was amplified by nested polymerase chain reaction （nPCR）. DNA sequence was determined, genotyping was performed by BLAST similarity search, and multiple alignment was performed with CLUSTAL X. Then a phylogenetie tree was constructed by Mega 3 software to illustrate the evolutionary relationships between clinical isolates and reference strains. Ninetysix specimens were sequenced, and 28 genetic variants were detected, among which E was the most prevalent genotype. The ompl gene was highly conserved for genotypes E and F, but appeared slightly less conserved for other genotypes, where the sequences displayed one to several nueleotide substitutions relative to the reference sequence. Phylogenetie tree showed that C. traehomatis serotypes were mainly divided into three clusters, according to previous grouping in the B, F-G, and C complexes, and the clinical isolates were highly related to the corresponding reference strains. It concluded that the ompl gene of the isolated C. traehomatis strains exhibited remarkable DNA sequence polymorphism, which can encourage for vaccine design and infection control.
