An investigation of alternative phosphorylation sites in arsR in Helicobacter pylori(part1)

Over last summer, I conducted a series of experiments to test out my hypothesis: 47th and 59th positions are alternative phosphorylation sites for arsR in H.pylori with the presence of glutamic acid at 52nd position.

I created mutants through mutagenesis, prepared mutant plasmids and did natural transformation on E.Coli. Sanger dideoxy DNA sequencing confirmed the alanine substitution at positions 47 and 59, with glutamic acid substitution at 52nd position. Then I incubated control strain, D52E mutant strain, and D47A 52E 59A mutant strain in pH5 and pH7 conditions respectively. I did mRNA extraction followed by cDNA synthesis to create the complementary DNA needed for qRT-PCR.

 

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