for the expression of adhesins of helicobacter pylori, I need to make a choice of housekeeping gene but I do not know which to choose.
Helicobacter pylori (H.pylori) has unipolar flagella made up of FlaA and FlaB proteins encoded by flaA and flaB genes. These regions are conserved and can be found in all strains of H. pylori. In this study based on bioinformatics’ methods, flaA and flaB nucleotide sequences of forty H.pylori complete genomes were extracted from NCBI databank and aligned with MEGA4 software. Based on conserved regions of flaA and flaB genes, 6 primer pairs were designed by AlleleID6 software. The designed primers were compared with glmM primers. Polymerase chain reactions (PCR) were performed with six primer pairs, twenty clinical samples and H. pylori 26695 as positive control. Primer Blast showed that three pairs of our designed primers were able to detect all 40 standard strains while glmM primers used in many papers were not capable of detecting all 40 standard strains of H. pylori. Although, few missense and some silent mutations, in comparison with H. pylori26695, were identified in flaA and flaB genes in this study, there was more semblance between them and other standard strains. So, it would be better to use these markers instead of glmM primers for detection of H.pylori as diagnostic kits.
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