Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats

Pseudomonas aeruginosa is a non-sporulating Gram negative, aerobic bacillus universally distributed in natural environment such as soil and water, which causes serious lethal infections. Its main targets are immunocompromised patients with attenuated host defense function which include burn victims,...

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Main Author:	Md. Safiul Alam Bhuiyan
Format:	Thesis
Language:	English English
Published:	2015
Subjects:	QR1-502 Microbiology
Online Access:	https://eprints.ums.edu.my/id/eprint/38613/1/24%20PAGES.pdf https://eprints.ums.edu.my/id/eprint/38613/2/FULLTEXT.pdf
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spelling	my-ums-ep.386132024-05-02T08:51:52Z Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats 2015 Md. Safiul Alam Bhuiyan QR1-502 Microbiology Pseudomonas aeruginosa is a non-sporulating Gram negative, aerobic bacillus universally distributed in natural environment such as soil and water, which causes serious lethal infections. Its main targets are immunocompromised patients with attenuated host defense function which include burn victims, organ transplant and cancer patient's long term therapy involves the use of antibiotics. Currently, there is no commercially available vaccine that can confer immunity to P. aeruginosa; therefore this study was conducted to determine the antigenic potential of a protein associated with the cell surface, which could subsequently be translated into a DNA based vaccine. The complete genome of P. aeruginosa strain PA0l was screened to determine an outer membrane surface coat Type III secretion protein (pscC) which is an important virulence determinant of type III secretion system (T3SS). Specific primers used were designed based on P. aeruginosa PA0l genome sequence available at the GenBank (NP250407) and subsequently cloned into two different Escherichia coli expression vector pGS-21a and pET-22b. The recombinant protein was analyzed by MALDI TOF-TOF mass spectrometer followed by purification using size exclusion chromatography. Finally, pscC gene was cloned onto a mammalian expression vector for plasmid immunization. The pMC2. l-pscC recombinant plasmid was directly injected intramuscularly in laboratory Sprague Dawley rats. Recombinant pscC antigen induced a specific humoral immune response against the antigen which was validated by agglutination and ELISA tests. The results clearly demonstrated that anti-pscC antibody was elicited using the animal model. The antibody level increased in 3 weeks of post immunization of all experimental doses compared with control group. The surface virulence Type III secretion protein (pscC) which is encoded by the outer membrane of T3SS genes will lead to the development of commercial plasmid vaccines to induce protective immunity against virulent Pseudomonas infection. 2015 Thesis https://eprints.ums.edu.my/id/eprint/38613/ https://eprints.ums.edu.my/id/eprint/38613/1/24%20PAGES.pdf text en public https://eprints.ums.edu.my/id/eprint/38613/2/FULLTEXT.pdf text en validuser masters Universiti Malaysia Sabah Institut Penyelidikan Bioteknologi
institution	Universiti Malaysia Sabah
collection	UMS Institutional Repository
language	English English
topic	QR1-502 Microbiology
spellingShingle	QR1-502 Microbiology Md. Safiul Alam Bhuiyan Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats
description	Pseudomonas aeruginosa is a non-sporulating Gram negative, aerobic bacillus universally distributed in natural environment such as soil and water, which causes serious lethal infections. Its main targets are immunocompromised patients with attenuated host defense function which include burn victims, organ transplant and cancer patient's long term therapy involves the use of antibiotics. Currently, there is no commercially available vaccine that can confer immunity to P. aeruginosa; therefore this study was conducted to determine the antigenic potential of a protein associated with the cell surface, which could subsequently be translated into a DNA based vaccine. The complete genome of P. aeruginosa strain PA0l was screened to determine an outer membrane surface coat Type III secretion protein (pscC) which is an important virulence determinant of type III secretion system (T3SS). Specific primers used were designed based on P. aeruginosa PA0l genome sequence available at the GenBank (NP250407) and subsequently cloned into two different Escherichia coli expression vector pGS-21a and pET-22b. The recombinant protein was analyzed by MALDI TOF-TOF mass spectrometer followed by purification using size exclusion chromatography. Finally, pscC gene was cloned onto a mammalian expression vector for plasmid immunization. The pMC2. l-pscC recombinant plasmid was directly injected intramuscularly in laboratory Sprague Dawley rats. Recombinant pscC antigen induced a specific humoral immune response against the antigen which was validated by agglutination and ELISA tests. The results clearly demonstrated that anti-pscC antibody was elicited using the animal model. The antibody level increased in 3 weeks of post immunization of all experimental doses compared with control group. The surface virulence Type III secretion protein (pscC) which is encoded by the outer membrane of T3SS genes will lead to the development of commercial plasmid vaccines to induce protective immunity against virulent Pseudomonas infection.
format	Thesis
qualification_level	Master's degree
author	Md. Safiul Alam Bhuiyan
author_facet	Md. Safiul Alam Bhuiyan
author_sort	Md. Safiul Alam Bhuiyan
title	Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats
title_short	Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats
title_full	Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats
title_fullStr	Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats
title_full_unstemmed	Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats
title_sort	evaluate the immunogenicity of plasmid encoding pscc protein of pseudomonas aeruginosa in rats
granting_institution	Universiti Malaysia Sabah
granting_department	Institut Penyelidikan Bioteknologi
publishDate	2015
url	https://eprints.ums.edu.my/id/eprint/38613/1/24%20PAGES.pdf https://eprints.ums.edu.my/id/eprint/38613/2/FULLTEXT.pdf
_version_	1804890330822082560

Evaluate the immunogenicity of plasmid encoding pscC protein of Pseudomonas aeruginosa in rats

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