THE EFFECT OF H. PYLORI PROTEINS JHP 0290 (FL) AND JHP 0290 ON HOST CELL BEHAVIOR
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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
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