Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats

Proteins associated with the bacterial membrane can be recruited for application as antigens for the development of vaccines. This preliminary study was directed towards evaluating the antigenic properties of the Pseudomonas aeruginosa (PA01) pscC protein which is a component of the Type III secreti...

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Main Authors: Md Safiul Alam Bhuiyan, Daisy Vanitha John, Fernades Opook, Hafiza Sultana, Kenneth Francis Rodrigues
Format: Article
Language:English
Published: Open Science Publishers 2017
Online Access:https://eprints.ums.edu.my/id/eprint/19079/1/Characterization%20of%20the%20pscC.pdf
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author Md Safiul Alam Bhuiyan
Daisy Vanitha John
Fernades Opook
Hafiza Sultana
Kenneth Francis Rodrigues
author_facet Md Safiul Alam Bhuiyan
Daisy Vanitha John
Fernades Opook
Hafiza Sultana
Kenneth Francis Rodrigues
author_sort Md Safiul Alam Bhuiyan
collection UMS
description Proteins associated with the bacterial membrane can be recruited for application as antigens for the development of vaccines. This preliminary study was directed towards evaluating the antigenic properties of the Pseudomonas aeruginosa (PA01) pscC protein which is a component of the Type III secretion system. Gene specific primers were designed to isolate the pscC gene which was isolated, ligated onto the multiple cloning site of vector pGS21(a), cloned and expressed in Escherichia coli (BL21). The molecular weight of the expressed pscC protein was determined by SDS-PAGE (10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis) and was found to be around 57 KDa and purified by the size exclusion chromatography. Finally, the purified pscC protein was injected subcutaneously into adult Sprague Dawley rats with a range of concentrations (50, 100 and 150 microgram per rat) respectively. Recombinant pscC antigen induced a specific humoral immune response against the antigen, which was validated by Enzyme-linked immunosorbent assay (ELISA). The results concluded that anti-pscC antibody was elicited in the animal model.
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spelling ums.eprints-190792018-03-06T00:40:47Z https://eprints.ums.edu.my/id/eprint/19079/ Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats Md Safiul Alam Bhuiyan Daisy Vanitha John Fernades Opook Hafiza Sultana Kenneth Francis Rodrigues Proteins associated with the bacterial membrane can be recruited for application as antigens for the development of vaccines. This preliminary study was directed towards evaluating the antigenic properties of the Pseudomonas aeruginosa (PA01) pscC protein which is a component of the Type III secretion system. Gene specific primers were designed to isolate the pscC gene which was isolated, ligated onto the multiple cloning site of vector pGS21(a), cloned and expressed in Escherichia coli (BL21). The molecular weight of the expressed pscC protein was determined by SDS-PAGE (10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis) and was found to be around 57 KDa and purified by the size exclusion chromatography. Finally, the purified pscC protein was injected subcutaneously into adult Sprague Dawley rats with a range of concentrations (50, 100 and 150 microgram per rat) respectively. Recombinant pscC antigen induced a specific humoral immune response against the antigen, which was validated by Enzyme-linked immunosorbent assay (ELISA). The results concluded that anti-pscC antibody was elicited in the animal model. Open Science Publishers 2017-01 Article PeerReviewed text en https://eprints.ums.edu.my/id/eprint/19079/1/Characterization%20of%20the%20pscC.pdf Md Safiul Alam Bhuiyan and Daisy Vanitha John and Fernades Opook and Hafiza Sultana and Kenneth Francis Rodrigues (2017) Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats. Journal of Applied Biology & Biotechnology, 5 (1). 035-041. ISSN 2347-212X http://doi.org/10.7324/JABB.2017.50106
spellingShingle Md Safiul Alam Bhuiyan
Daisy Vanitha John
Fernades Opook
Hafiza Sultana
Kenneth Francis Rodrigues
Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats
title Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats
title_full Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats
title_fullStr Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats
title_full_unstemmed Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats
title_short Characterization of the pscC (Type III secretion) gene of Pseudomonas aeruginosa (PA01) and assessment of immunogenicity of pscC protein in rats
title_sort characterization of the pscc type iii secretion gene of pseudomonas aeruginosa pa01 and assessment of immunogenicity of pscc protein in rats
url https://eprints.ums.edu.my/id/eprint/19079/1/Characterization%20of%20the%20pscC.pdf
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