THE EFFECT OF H. PYLORI PROTEINS JHP 0290 (FL) AND JHP 0290 ON HOST CELL BEHAVIOR
Article Sidebar
Main Article Content
Abstract
Objective: This study was designed to investigate the effect of H. Pylori proteins JHP 0290 (FL) and JHP 0290 on host cell response.
Materials and Methods: This study was conducted at Khyber Teaching Hospital (KTH), Peshawar from April 2021 to September 2021. H. Pylori genomic DNA (J99 and 26695), pET expression system plasmid (PET21a, 22b & 28b+), kanamycin 50µg/ml and ampicillin 100µg/ml, LB agar plates and LB media, Talon@IMAC resin column were used for studying H. pylori proteins. Genes coding for these proteins were successively cloned into PET21a, PET22b, and PET28b+ expression vectors but no expression was seen with different concentrations of IPTG in BL21(DE3) culture, and expression was prolonged for 4hrs.
Results: Only JHP0290 (FL) & JHP0290 (-17a.a) were successively cloned in PET28b+ and expression was induced by using 100µM IPTG in E.coli strain (BL21DE3). Restriction analysis showed the size of JHP0290 (FL & -17a.a) at approximately 550bp and protein size was analyzed on SDS-page which was 20Kda approximately. JHP0290FL protein was found in pellets while JHP0290 (-17a.a) protein was found in pellets & supernatant before being applied to Talon cell through resin column to get purified protein. The study showed a clear band of JHP0290 (FL) on SDS-page with imidazole eluted fraction pooled, while a faint band with a distinct band of JHP0290 (-17a.a) was also seen on SDS-page.
Conclusion: These changes are indicative of altered host cell response induced by H.pylori protein JHP0290 suggesting its possible involvement in the development and severity of the disease.
Keywords: H.pylori proteins, virulence factor, host cell response, genomic DNA, pathogenicity
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
All articles published in the Journal of Medical Sciences (JMS) are licensed under the Creative Commons Attribution 4.0 International License (CC-BY 4.0). Under the CC BY 4.0 license, author(s) retain the ownership of the copyright publishing rights without restrictions for their content, and allow others to copy, use, print, share, modify, and distribute the content of the article even for commercial purposes as long as the original authors and the journal are properly cited. No permission is required from the author/s or the publishers for this purpose. Appropriate attribution can be provided by simply citing the original article. The corresponding author has the right to grant on behalf of all authors, a worldwide license to JMS and its licensees in all forms, formats, and media (whether known now or created in the future), The corresponding author must certify and warrant the authorship and proprietorship and should declare that he/she has not granted or assigned any of the article’s rights to any other person or body.
The corresponding author must compensate the journal for any costs, expenses, or damages that the JMS may incur as a result of any breach of these warranties including any intentional or unintentional errors, omissions, copyright issues, or plagiarism. The editorial office must be notified upon submission if an article contains materials like text, pictures, tables, or graphs from other copyrighted sources. The JMS reserves the right to remove any images, figures, tables, or other content, from any article, whether before or after publication, if concerns are raised about copyright, license, or permissions and the authors are unable to provide documentation confirming that appropriate permissions were obtained for publication of the content in question.
