Construction and identification of Chinese mutant <i>Helicobacter pylori</i> strain with absent expression of cagA gene

Huang, Zhi-Gang; Duan, Guangcai; Fan, Qi-Wen; Huang, Xueyong

doi:10.11569/wcjd.v14.i33.3190

Cited by 1 publication

(1 citation statement)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In brief, we produced a Hp27_∆ cagA isogenic mutant harboring a total cagA deletion as previously described [13] . H pylori were grown on blood agar plates under microaerophilic conditions for 3 d and genomic DNA was extracted with a genomic DNA purification kit (Takara).…”

Section: Construction Of Hp27_∆caga Isogenic Mutantmentioning

confidence: 99%

Mutation of cytotoxin-associated gene A affects expressions of antioxidant proteins of Helicobacter pylori

Huang¹,

Duan²,

Fan³

et al. 2009

WJG

Self Cite

View full text Add to dashboard Cite

AIM:To determine if disruption of the cagA gene of Helicobacter pylori (H pylori ) has an effect on the expression of other proteins at proteome level.METHODS: C o n s t r u c t i o n o f a cagA k n o c k o u t m u t a n t H p 2 7 _ ∆c a g A (c a g A -) v i a h o m o l o g o u s re c o m b i n a t i o n w i t h t h e w i l d -ty p e s t ra i n Hp27 (cagA + ) as a recipient was performed. The method o f s o n i c a t i o n -u r e a -C H A P S -DTT wa s e m p l o ye d to extract bacterial proteins from both strains. Soluble proteins were analyzed by two-dimensional electrophoresis (2-DE). Images of 2-DE gels were digitalized and analyzed. Only spots that had a statistical significance in differential expression were selected and analyzed by matrix-assisted laser desorption/ionizationtime of flight mass spectrometry (MALDI-TOF-MS). Biological information was used to search protein database and identify the biological function of proteins. RESULTS:The proteome expressions between wild-type strain and isogenic mutant with the cagA gene knocked-out were compared. Five protein spots with high abundance in bacteria proteins of wild-type strains, down-regulated or absently expressed in bacteria proteins of mutants, were identified and analyzed. From a quantitative point of view, the identified proteins are related to the cagA gene and important antioxidant proteins of H pylori , including alkyl hydroperoxide reductase (Ahp), superoxide dismutase (SOD) and modulator of drug activity (Mda66), respectively, suggesting that cagA is important to maintain the normal activity of antioxidative stress and ensure H pylori persistent colonization in the host. C O N C L U S I O N :c a g A g e n e i s r e l e v a n t t o t h e expressions of antioxidant proteins of H pylori , which may be a novel mechanism involved in H pylori cagA pathogenesis.

show abstract

Section: Construction Of Hp27_∆caga Isogenic Mutantmentioning

confidence: 99%