Cloning of urease operon from helicobacter pylori J99

Abstract

Helicobacter pylori is a gram-negative bacterium that colonizes the gastric mucosal in human stomach. It causes gastric ulcers and cancer by mechanisms that are not well understood. Helicobacter pylori produces enzyme that damage gastric epithelial cells, such as urease (constitute up to 10% of total bacterial protein) that neutralize the acidic medium permitting its survival in the human’s stomach. Urease (urea amidohydrolase; EC 3.5.1.5) is a nickel-dependent enzyme that catalyzes the hydrolysis of urea to form ammonia and carbon dioxide. The genes for urease enzymes are organized in a gene cluster or operon. In this preliminary work, the urease operon was amplified using polymerase chain reaction (PCR) and cloned into pET 32Ek/LIC (Invitrogen, USA) system. The aim of this study is to obtain a functional replicon of urease operon that capable of expressing the urease enzymes and introduce the replicon in animal model for pathogenesis, as well as, pathophysiological studies of Helicobacter pylori.