Analysis of 16S rRNA by PCR, RFLP and RAPD fingerprinting of different <i>Helicobacter</i> species.

<p>Panel A: DNA isolated from various <i>Helicobacter</i> species including the Bengal tiger isolate was subjected to 16SrRNA PCR. A conserved 1.5 kb DNA fragment of the genus <i>Helicobacter</i> was amplified <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071220#pone.0071220-Terio1" target="_blank">[52]</a>. Panel B: To confirm their specificity, all PCR products were then investigated by RFLP (restriction fragment length polymorphism) using the restriction endonuclease <i>Alu</i>I. The <i>Alu</i>I pattern of the tiger isolate gave rise to 3 major bands (asterisks) which were unique to all other <i>Helicobacters</i>. Some similar bands were only found in the RFLP pattern of <i>H. pylori</i> and <i>H. acinonychis</i> (asterisks), indicating their close genetic relatedness. Panel C: RAPD fingerprinting of the <i>Helicobacter</i> isolates using primer D9355 was performed as described previously <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071220#pone.0071220-Akopyanz1" target="_blank">[27]</a>. A typical RAPD fingerprinting profile is shown and revealed the relatedness between <i>H. pylori</i>, <i>H. acinonychis</i> and Bengal tiger isolate. Asterisks indicate three major bands which were identical among the latter three samples. M, DNA size marker.