[Construction and identification of ureB gene vaccine of H. pylori].
OBJECTIVE
To clone ureB gene of H.pylori and construct its gene vaccine.


METHODS
ureB gene was amplified by PCR from genome of H. pylori 11637 strain and subcloned into pMD18-T vector. The vector digested with restriction enzyme (Sal I and Bgl II) was inserted into pTCAE and transformed into E. coli DH5alpha. The positive recombinant plasmid identified by digesting with restriction enzyme (Sal I and Xho I) and sequencing named pT-ureB. The pT-ureB was transfected into CHO cells by electroporation method. The expression of UreB protein was detected by Western blot.


RESULTS
The pT-ureB was obtained by cloning and recombinant DNA technique. The Western blot analysis showed that the expression of UreB protein (M(r)approximately 62,000) was detected in culture supernatant of CHO cells following transfection with pT-ureB.


CONCLUSIONS
UreB DNA vaccine of H. pylori was successfully constructed. The expression of UreB protein can be detected in culture supernatants of transfected CHO cells.
Zeng
Wei-kun
WK
Xinqiao Hospital, Third Military Medical University, Chongqing 400038, China.
Zou
Quan-ming
QM
Jing
Shen-rong
SR
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005822:Genetic Vectors / Q000235:genetics; D016480:Helicobacter pylori / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014510:Urease / Q000235:genetics* / Q000378:metabolism; D019444:Vaccines, DNA / Q000235:genetics*
D001426:Bacterial Proteins; D019444:Vaccines, DNA; D014510:Urease