幽门螺杆菌jhp947和cagA的检测及jhp947真核表达载体的构建

The relationship between jhp947 and cagA in gastroduodenal diseases and construction of a eukaryotic expression vector of jhp947

目的研究幽门螺杆菌(Helicobacter pylori,Hp)cagA和可塑区jhp947基因与胃及十二指肠疾病的关系;构建jhp947真核表达载体,为进一步研究jhp947的功能奠定基础。方法从消化性疾病患者胃粘膜活检组织中分离培养Hp,PCR分离cagA和jhp947基因,分析基因与疾病的关系;从Hp国际标准株J99中获取jhp947全长基因,克隆至pcDNA3.1/myc-HisA载体并鉴定。结果分离获得108株Hp,目的基因在胃炎、溃疡、胃癌组织中的检出率分别为:jhp9473.2%、32.2%和33.3%,cagA74.2%、93.5%和86.7%,jhp947检出率胃癌、溃疡组与胃炎组比较差异有统计学意义(χ2值分别为12.04和8.64,P均<0.01)。jhp947和cagA的检出率无显著相关性(P>0.05)。经鉴定jhp947真核表达载体构建正确。结论jhp947与胃癌和消化性溃疡的发生密切相关,jhp947和cagA在致病过程中可能没有协同作用,成功构建jhp947真核表达载体。

Objective To analyze the relationship between the genes jhp947 and cagA in Helicobacter pylori （Hp） in gastroduodenal diseases and to construct a eukaryotic expression vector of jhp947 of strain HpJ99. Methods Hp strains were isolated from gastric biopsy samples of patients with gastritis, peptic ulcers （PU）, or gastric carcinoma （GC） . A PCR-based methodology was used to detect jhp947 and cagA. Experimental data were analyzed by the χ^2 test using the partition of χ^2 method. Jhp947 was amplified by PCR and cloned into vector pcDNA3.1/myc-His A. Results Positive rates of jhp947 and cagA detected in gastritis, peptic ulcer, and gastric carcinoma samples were 73.2%, 32.2%, and 33.3% for jhp947 and 74.2%, 93.5%, and 86.7% for cagA, respectively. Positive rates of jhp947 detected in gastric carcinoma and peptic ulcer samples were higher than in gastritis samples （χ^2 = 12.04, P〈0.01;χ^2 =8. 64, P〈0.01）. Positive rates of jhp947 with positive and negative rates of cagA were 17.0% （15/88） and 10.0 % （2/20）, respectively （P〈0.05）. The eukaryotic expression vector of pcDNA3.1/myc-His A-947 was successfully constructed. Conclusion The jhp947 gene might be implicated in the development of both duodenal ulcers and gastric carcinoma. The presence of the jhp947 gene was not associated with the cagA-positive genotype. A eukaryotic expression vector of jhp947 was successfully constructed.