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Gastric Mucosa Quantitative Polymerase Chain Reaction Analysis for Detecting Helicobacter pylori and Antibiotic Resistance
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[Cloning and sequence analysis of UreB of Helicobacter pylori isolated from children]

Zhen-Wen Zhou1, Qiu-Lian Deng, Hui-Min Xia

  • 1Guangzhou Women and Children's Medical Center, Guangzhou 510120, China. zhouzhenwen28@126.com

Zhongguo Dang Dai Er Ke Za Zhi = Chinese Journal of Contemporary Pediatrics
|February 2, 2010
PubMed

Insights

The UreB gene from a children's Helicobacter pylori strain was cloned into a pGEX-4T-1 plasmid. This successful cloning and sequence analysis of the UreB gene provides a foundation for developing oral H. pylori vaccines.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Helicobacter pylori (H. pylori) is a significant pathogen.
  • The UreB gene is a key component of H. pylori.
  • Understanding H. pylori genetics is crucial for vaccine development.

Purpose of the Study:

  • To clone the UreB gene from H. pylori strains isolated from children into the pGEX-4T-1 expression plasmid.
  • To perform sequence analysis of the cloned UreB gene.
  • To lay the groundwork for research into oral H. pylori vaccines.

Main Methods:

  • Designing specific primers for the H. pylori UreB gene.
  • Amplifying the UreB gene using Polymerase Chain Reaction (PCR) from clinical isolates.
  • Cloning the amplified UreB gene into the pGEX-4T-1 plasmid after enzymatic digestion.
  • Transforming the recombinant plasmid into E. coli BL21.
  • Identifying the recombinant plasmid via double enzyme digestion and DNA sequencing.

Main Results:

  • Successfully amplified and cloned the UreB gene (1710 bp) from H. pylori strain GZCH1.
  • Confirmed the correct open reading frame of the UreB gene through DNA sequencing.
  • Sequence analysis revealed 98% identity at both DNA and amino acid levels with other H. pylori strains.
  • The UreB gene sequence was submitted to GenBank (accession number: FJ455126).

Conclusions:

  • The UreB gene of H. pylori strain GZCH1 was successfully cloned into the pGEX-4T-1 expression vector.
  • This cloning provides a crucial basis for further research on oral H. pylori vaccines.
  • The high sequence identity suggests conserved UreB function across different H. pylori strains.
Abstract

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