人幽门螺杆菌外膜蛋白oipA编码基因的克隆与序列分析
Cloning and sequence analysis of gene oipA encoding an outer membrane protein of human Helicobacter pylori.
作者信息
Chen Dao-Rong, Huang Ai-Long, Tao Xiao-Hong, Wang Pi-Long, Jiang Zheng
机构信息
Department of Gastroenterology, the First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400016, China.
出版信息
World J Gastroenterol. 2004 Nov 1;10(21):3205-7. doi: 10.3748/wjg.v10.i21.3205.
AIM
To construct a recombinant E. coli strain that would highly express the proinflammatory outer membrane protein of human Helicobacter pylori (H pylori).
METHODS
The oipA DNA was amplified by PCR, inserted into pET-32a, and transformed into Top10 E. coli strain. This recombinant plasmid of Top10 was sent out for nucleotide sequence analysis. Finally this sequence AF479754 was compared with HP0638 and JHP0581.
RESULTS
The sequence of the aim gene was obtained. It had 924 base pairs. The identity was 95.32% against HP0638, 95.02% against JHP0581, which was higher than the identity between HP0638 and JHP0581.
CONCLUSION
Although the aim gene was obtained, but it was different from the published sequence of GenBank. It is not clear what makes this difference. Maybe it is because different strain was used or because there were some variations. So more researches are required to prove it.
目的
构建一种能高效表达人幽门螺杆菌（H pylori）促炎外膜蛋白的重组大肠杆菌菌株。
方法
通过聚合酶链反应（PCR）扩增oipA DNA，将其插入pET - 32a中，并转化到Top10大肠杆菌菌株中。将Top10的这种重组质粒送去进行核苷酸序列分析。最后将该序列AF479754与HP0638和JHP0581进行比较。
结果
获得了目的基因的序列。它有924个碱基对。与HP0638的同一性为95.32%，与JHP0581的同一性为95.02%，高于HP0638和JHP0581之间的同一性。
结论
虽然获得了目的基因，但它与GenBank公布的序列不同。尚不清楚造成这种差异的原因。可能是因为使用了不同的菌株，或者存在一些变异。所以需要更多研究来证实这一点。
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