Shi Yan-yan, Ding Shi-gang, Lu Feng-min, Zhang Jing, Chen Xiang-mei, Liu Lin-na, Wang Ye
Department of Gastroenterology, Peking University Third Hospital, Beijing 100191, China.
Zhonghua Yi Xue Za Zhi. 2010 Nov 2;90(40):2830-3.
To analyze the gene sequences of Trx1 and Trx2 of Helicobacter pylori (HP) from different gastric diseases, define the amino acid sequences and investigate the relationship with these diseases.
The HP strains were isolated from gastric mucosa of 25 patients with chronic gastritis, peptic ulcer and gastric cancer respectively and cultured on solid blood agar medium. The primers of Trx1 and Trx2 were designed according to the sequences of GenBank. The Trx1 and Trx2 were respectively amplified by PCR (polymerase chain reaction) and were sequenced by the bioinformatics method. The sequences were compared with those of the international standard HP strains.
The whole sequence of Trx1 and the first 295 bp of Trx2 were successfully amplified to allow for sequence comparison by BioEdit. The Trx1 from different HP strains contained 321 bp encoding 106 amino acids. The mutation ratio was 13.4% (43/321). All amino acids contained the same active motif Cys-Gly-Pro-Cys. The homology of Trx1 amino acids from 25 strains was 97.2% (103/106). The mutation ratio for first 295 bp of Trx2 was 18.6% (55/295). All amino acids encoded by Trx2 contained one CXXC zone while the Cys-Gly-Pro-Cys motif was not found. The homology of Trx2 amino acids was 84.7% (83/98).
Two different subtypes of Trx from clinically isolated Hp strains have mutation sites. But the homology of encoded amino acids is relatively high because of invalid mutations. Trx1 of HP strains from different diseases all contains a redox active motif Cys-Gly-Pro-Cys while Trx2 contains only a CXXC zone. The above results will provide valuable rationales for future studies of the biological function and pathogenic mechanisms of HP Trx1 and Trx2.
分析不同胃部疾病中幽门螺杆菌(HP)的Trx1和Trx2基因序列,确定氨基酸序列,并研究其与这些疾病的关系。
分别从25例慢性胃炎、消化性溃疡和胃癌患者的胃黏膜中分离出HP菌株,并在固体血琼脂培养基上培养。根据GenBank序列设计Trx1和Trx2的引物。通过聚合酶链反应(PCR)分别扩增Trx1和Trx2,并采用生物信息学方法进行测序。将序列与国际标准HP菌株的序列进行比较。
成功扩增出Trx1的全序列和Trx2的前295 bp,以便通过BioEdit进行序列比较。不同HP菌株的Trx1包含321 bp,编码106个氨基酸。突变率为13.4%(43/321)。所有氨基酸均含有相同的活性基序Cys-Gly-Pro-Cys。25株Trx1氨基酸的同源性为97.2%(103/106)。Trx2前295 bp的突变率为18.6%(55/295)。Trx2编码的所有氨基酸均含有一个CXXC区域,而未发现Cys-Gly-Pro-Cys基序。Trx2氨基酸的同源性为84.7%(83/98)。
临床分离的Hp菌株的两种不同亚型的Trx存在突变位点。但由于无效突变,编码氨基酸的同源性相对较高。不同疾病的HP菌株的Trx1均含有氧化还原活性基序Cys-Gly-Pro-Cys,而Trx2仅含有一个CXXC区域。上述结果将为今后研究HP Trx1和Trx2的生物学功能和致病机制提供有价值的理论依据。