Construction of a Plasmid Vector for Expression of Bacteriocin N15-encoding Gene and Effect of Engineered Bacteria on Enterococcus Faecalis
نویسندگان
چکیده
A 6.09-kb plasmids vector pOri253 was constructed from the plasmid pIL253 (5.2 kb) and a 0.89-kb fragment of oriColE1 from pBluescript II KS. The bifunctional plasmid pOri253 conferred erythromycin resistance in both Escherichia coli and Enterococcus faecalis. It has a unique site for each of EcoRI, BamHI, SalI and PstI derived from pIL253. The plasmid is quite stable in E. faecalis JCM8726 when cultured in Mann Rogosa Sharpe broth without antibiotic. The lactococcal promoter P23 was inserted at one end of the pOri253 multicloning site. Gene expression was assessed by using an entAI gene which coded for bacteriocin N15. The E. faecalis harboring constructed plasmid carrying P23 (pOrient23) had more antibacterial activity than parental E. faecalis JCM8726 or its clone harboring non-P23 containing plasmid (pOrient), as determined by means of an overlay method.
منابع مشابه
Construction of an Expression Plasmid (Vector) Encoding Brucella melitensis Outer Membrane Protein, a Candidate for DNA Vaccine
Background: DNA vaccination with plasmid encoding bacterial, viral, and parasitic immunogens has been shown to be an attractive method to induce efficient immune responses. Bacteria of the genus Brucella are facultative intracellular pathogens for which new and efficient vaccines are needed. Methods: To evaluate the use of a DNA immunization strategy for protection against brucellosis, a pla...
متن کاملEngineered and construction of pDS132::∆virG as suicide vector for targeted gene deletion of virG from Shigella flexneri 2a in order to generation a live attenuated Shigella vaccine
Background & Objective: Shigella are Gram negative bacteria capable of inducing their entry into non-phagocytic cells via secretion of various effector proteins called invasion plasmid antigens (Ipas). The most important of them is VirG protein. Live attenuated Shigella vaccines have indicated promise in inducing protective immune responses in human clinical trials. In current situation, const...
متن کاملExpression of Brucella abortus Omp25 Protein in Lactococcus lactis Probiotic Bacteria
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. Materi...
متن کاملHeterologous extracellular production of enterocin P from Enterococcus faecium P13 in the methylotrophic bacterium Methylobacterium extorquens.
Enterocin P (EntP), a strong antilisterial pediocin-like bacteriocin from Enterococcus faecium P13, was produced by Methylobacterium extorquens. For heterologous expression of EntP in the methylotrophic bacterium M. extorquens, a recombinant plasmid was constructed. The gene encoding the EntP structural gene (entP) was cloned into the plasmid vector pCM80, under control of the methanol dehydrog...
متن کاملCharacterization and cloning of the genes encoding enterocin 1071A and enterocin 1071B, two antimicrobial peptides produced by Enterococcus faecalis BFE 1071.
The pH-neutral cell supernatant of Enterococcus faecalis BFE 1071, isolated from the feces of minipigs in Göttingen, inhibited the growth of Enterococcus spp. and a few other gram-positive bacteria. Ammonium sulfate precipitation and cation-exchange chromatography of the cell supernatant, followed by mass spectrometry analysis, yielded two bacteriocin-like peptides of similar molecular mass: en...
متن کامل