Cloning and sequence analysis of UreB of Helicobacter pylori isolated from children

ZHOU Zhen-Wen; DENG Qiu-Lian; XIA Hui-Min; GENG Lan-Lan; LIANG Wei-He; XIE Yong-Qiang; HUANG Yong; GONG Si-Tang

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Chinese Journal of Contemporary Pediatrics ›› 2009, Vol. 11 ›› Issue (12) : 877-880.

CLINICAL RESEARCH

Cloning and sequence analysis of UreB of Helicobacter pylori isolated from children

ZHOU Zhen-Wen, DENG Qiu-Lian, XIA Hui-Min, GENG Lan-Lan, LIANG Wei-He, XIE Yong-Qiang, HUANG Yong, GONG Si-Tang

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Abstract

OBJECTIVE: To clone UreB gene of Helicobacter pylori (H. pylori) isolated from children to pGEX-4T-1 expression plasmid, and do sequence analysis. METHODS: A pair of specific primer was designed according to H. pylori UreB gene in the GenBank. Using H. pylori strains isolated from children as a template, a UreB gene was obtained by PCR. After EcoR I and Not I digestion, the PCR production was linked with pGEX-4T-1 which was digested with the same enzymes. The recombinant plasmid was transformed into E.coli BL21 and identified by double enzyme digestion and sequence analysis. The sequence results were compared with the gene sequence in the GenBank. RESULTS: A UreB gene was successfully amplified from children′s H. pylori strain GZCH1. It was 1 710 bp in size. The objective band was identified by double enzyme digestion. DNA sequence showed that UreB was in the correct open reading frame. The sequence comparison analysis showed that DNA and amino acid sequence identities of UreB gene with other strains were 98%. The sequence of UreB of H. pylori strain GZCH1 was submitted to GenBank (accession number：FJ455126). CONCLUSIONS: UreB of H. pylori strain GZCH1 is successfully cloned to pGEX-4T-1, which provides a basis for research of oral H. pylori vaccine.［Chin J Contemp Pediatr, 2009, 11 (11):877-880］

Key words

Helicobacter pylori / UreB / Clone / Child

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ZHOU Zhen-Wen, DENG Qiu-Lian, XIA Hui-Min, GENG Lan-Lan, LIANG Wei-He, XIE Yong-Qiang, HUANG Yong, GONG Si-Tang. Cloning and sequence analysis of UreB of Helicobacter pylori isolated from children[J]. Chinese Journal of Contemporary Pediatrics. 2009, 11(12): 877-880

References

［1］Warren J, Marshall B. Unidentified curved bacilli on gastric epithelium inactive chronic gastritis［J］. Lacent, 1983, 1(8336):1273-1275.
［2］Stephenie CS, Alfonso JB. Helicobacter pylori strikes again-gastric mucosa-associated lymphoid tissue(MALT) lymphoma［J］. Gastroenterol Nurs, 2007, 30(5):348-354.
［3］Danesh J. Helicobacter pylori infection and gastric cancer: systematic review of epidemiological studies［J］. Aliment Pharmacol Ther, 1999, 13(7):851-856.
［4］王娟,王润.中国幽门螺杆菌感染流行病学杂志Mata分析［J］.中华流行病学杂志, 2003, 24(6):443-446.
［5］耿岚岚,龚四堂,区文玑,潘瑞芳,陈宝心,丘小汕.两种非侵入性诊断方法在儿童幽门螺杆菌感染中的应用［J］.实用儿科临床杂志, 2003, 18(7):538-539.
［6］关克涛,徐文泉,刘云锋,卢铭江,颜幕霞.酶联免疫吸附测定检测儿童粪便幽门螺杆菌抗原［J］.广州医学院学报, 2006, 34(4):43-45.
［7］黄烈平,庄满利,顾承萍.36株青少年幽门螺杆菌耐药分析［J］.中国当代儿科杂志, 2009,11(3):210-212.
［8］袁小澎,邹全明,柏杨,杨珺,郭鹰,张卫军,等. 幽门螺杆菌B亚单位功能片段的纯化及活性研究［J］.南方医科大学学报, 2007, 27(7):959-962.
［9］赵煜,徐晓华,刘风霖,张书红,司徒爱明.儿童幽门螺杆菌感染与相关性疾病的临床研究［J］. 中国当代儿科杂志, 2008, 10(3):403-404.
［10］Myers GA, Ermak TH, Georgakopoulos K, Tibbitts T, Ingrassia J, Gray H, et al. Oral immunization with recombinant Helicobacter pylori urease confers long-lasting immunity against Helicobacter felis infection［J］.Vaccine, 1999, 17(12):1394-1403.
［11］Liu X, Hu J, Zhang X, Fan D. Oral immunization of mice with attenuated Salmonella typhimurium espressing Helicobacter pylori urease B subunit［J］. Chin Med J, 2002, 115(10):1513-1516.
［12］Michetti P, Kreiss C, Kotloff KL, Pota N, Bachmann D, Herranz M, et al. Oral immunization with urease and Escherichia coli heat-labile enterotoxin is safe and immunogenic in Heliobacter pylori-infected adults［J］. Gastroenterology, 1999, 116(4):804-812.
［13］周珍文,胡旭初,余新炳.枯草杆菌芽孢载体疫苗的研究及其在寄生虫病防治中的应用前景［J］.热带医学杂志, 2007, 7(9):935-937.
［14］Zhou ZW, Xia HM, Hu XC, Huang Y, Li YW, Li L, et al. Oral administration of a Bacillus subtilis Spore-based vaccine expressing Clonorchis sinensis tegumental protein 22.3 kDa confers protection against Clonorchis sinensis［J］.Vaccine, 2008, 26(15):1817-1825.
［15］Zhou ZW, Xia HM, HuXC, Huang Y, Ma CL, Chen XX, et al. Immunogenicity of recombinant Bacillus subtilis spores expressing Clonorchis sinesis tegumental protein［J］. Parasitol Res, 2008, 102(2):293-297.