[Purification and functional analysis of Helicobacter pylori UreB protein fragment].
OBJECTIVE
To establish an effective method for purification of Helicobacter pylori UreB fragment and conduct functional analysis of the purified protein.


METHODS
The protein fragment expression was induced by IPTG and the expressed protein was purified through affinity chromatography and ion-exchange chromatography. The purity of the fragment was determined by high-performance liquid chromatography (HPLC), and the specific biological activity of the purified fragment was assayed by urease activity inhibition test.


RESULTS
The protein fragment was highly expressed in E. coli with a purity over 91%. The protein fragment showed highly specific biological activity and the specific antibody induced by this fragment in rabbits could inhibit the activity of urease in a dose-dependent manner.


CONCLUSIONS
The UreB fragment with high purity and biological activity can be applied for further studies.
Yuan
Xiao-peng
XP
Department of Clinical Laboratory, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. yuanxp2001@yahoo.com.cn
Zhou
Quan-ming
QM
Bai
Yang
Y
Yang
Jun
J
Guo
Ying
Y
Zhang
Wei-jun
WJ
Liu
Zheng-xiang
ZX
D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000737:chemistry*; D001428:Bacterial Vaccines / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D004586:Electrophoresis; D004926:Escherichia coli / Q000235:genetics; D016480:Helicobacter pylori / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification*; D011817:Rabbits; D014510:Urease / Q000037:antagonists & inhibitors
D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D010446:Peptide Fragments; D014510:Urease