Construction of replicative and integrative plasmids for setting up the in vivo expression technology in Helicobacter pylori.
Some pathogenic factors of Helicobacter pylori, a bacterium involved in peptic ulcer and gastric cancer, have already been identified using either global or particular approach, but there are still some orphan genes and unidentified pathogenic factors. One of the methods used successfully for the identification of virulence genes of many pathogens is the in vivo expression technology. We describe here the construction and sequences of three different plasmids, one integrative and two replicatives, for the identification of virulence genes by using in vivo expression technology in H. pylori, and of potential use in other bacteria such as Campylobacter spp. Moreover, the use of the green fluorescent protein could allow to classify the genes according to the strength of their expression and to identify those which are repressed upon interaction with gastric mucosa.
Laboratoire de Bactériologie, Université Victor Segalen Bordeaux 2, Bordeaux, France.
Fabrice
F
Angelini
Armelle
A
Ménard
Corinne
C
Asencio
Armelle
A
Marais
Francis
F
Mégraud
D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005756:Gastritis / Q000382:microbiology*; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D016481:Helicobacter Infections / Q000382:microbiology*; D016480:Helicobacter pylori / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial; D014774:Virulence
D004269:DNA, Bacterial; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins