Expression of Brucella abortus Omp25 protein in Lactococcus lactis probiotic bacteria
Journal of Mazandaran University of Medical Sciences
; 32(217):16-31, 2023.
Article
in Persian
| CAB Abstracts | ID: covidwho-2295750
ABSTRACT
Background and purpose:
The sequence of Omp25 is conserved in all Brucella species. The high antigenicity of the product of this gene stimulates the host's immune system. Using engineered probiotic bacteria is an appropriate method for vaccine transport. The aim of this study was to express the Omp25 of the Brucella abortus pathogenic bacterium in Lactococcus lactis probiotic bacterium. Materials andmethods:
In this experimental study, the required vector was designed and synthesized to include the gene of interest and a signal peptide (pNZ8148-Usp45-Omp25). E. coli strain TOP10F was transformed using the pNZ8148-Usp45-Omp25 expression vector based on induction by nisin. The recombinant plasmid was extracted from the transformed bacteria using a plasmid extraction kit. The L. lactis was transformed by pNZ8148-Usp45-Omp25 vector using electroporation. Evaluation of the expression of Omp25 gene at the RNA level was assessed by reverse transcription method and confirming the presence of recombinant Omp25 protein in the engineered bacteria using SDS-PAGE method.Results:
Successful expression of B. abortus Omp25 in L. lactis was verified by RT-PCR. Subsequently, the proteins were separated based on molecular weight using sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). The protein expression analysis showed the expression of Omp25 as a 25 kDa extra band in transformed L. lactis compared to the L. lactis receiving the vector lacking the target gene.Conclusion:
This study shows that Omp25 is expressed in L. lactis transformed via pNZ8148-Usp45-Omp25 by electroporation. Transformed L. lactis can be successfully used as a subunit oral vaccine in prevention of Brucellosis.
Host Resistance and Immunity [HH600]; Animal Immunology [LL650]; Prion; Viral; Bacterial and Fungal Pathogens of Animals [LL821]; Genetic Engineering; Gene Transfer and Transgenics [WW100]; Diagnostic; Therapeutic and Pharmacological Biotechnology [WW700]; Molecular Biology and Molecular Genetics [ZZ360]; Genetics and Molecular Biology of Microorganisms [ZZ395]; bacterial proteins; brucellosis; gene expression; genes; genetic engineering; genetic vectors; nucleotide sequences; oral vaccination; plasmids; probiotics; recombinant vaccines; SDS-PAGE; strains; vaccination; vaccine development; vaccines; bacterial diseases; Brucella abortus; Lactococcus lactis; Sialodacryoadenitis virus; Brucella; Brucellaceae; Rhizobiales; Alphaproteobacteria; Proteobacteria; Bacteria; prokaryotes; Lactococcus; Streptococcaceae; Lactobacillales; Bacilli; Firmicutes; Rat coronavirus; Murine coronavirus; Betacoronavirus; Coronavirinae; Coronaviridae; Nidovirales; positive-sense ssRNA Viruses; ssRNA Viruses; RNA Viruses; viruses; undulant fever; genetic manipulation; cloning vectors; DNA sequences; sodium dodecyl sulfate-PAGE; bacterial infections; bacterioses; bacterium
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Collection:
Databases of international organizations
Database:
CAB Abstracts
Language:
Persian
Journal:
Journal of Mazandaran University of Medical Sciences
Year:
2023
Document Type:
Article
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