Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis.
Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis.
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应用FLP-FRT系统构建幽门螺杆菌无标记缺失及hp0788基因在发病机制中的功能研究
DOI：
10.3389/fmicb.2017.02357
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发表时间：
2017
影响因子：
5.2
通讯作者：
Zhao H
中科院分区：
文献类型：
--
作者：
Ji X;Wang Y;Li J;Rong Q;Chen X;Zhang Y;Liu X;Li B;Zhao H
幽门螺杆菌是一种革兰氏阴性、微需氧的细菌，与人类胃部疾病有关。为阐明幽门螺杆菌的致病机制以及感染的异质性问题，仍需对毒力基因进行进一步研究。为了开发一种高效、准确的方法来研究幽门螺杆菌致病过程中的基因功能，基于FLP - FRT重组系统建立了一种针对单个基因和大片段的无痕缺失方法。利用该方法，缺失了编码外膜蛋白（HofF）的基因hp0788。hp0788的缺失不影响幽门螺杆菌的生长和运动能力，但降低了细菌对胃上皮细胞的黏附能力。hp0788的缺失也减少了幽门螺杆菌感染导致的GES - 1细胞凋亡。这些结果表明hp0788参与了细菌 - 宿主相互作用，在幽门螺杆菌感染中起重要作用。此外，还利用FLP - FRT方法成功实现了在cag致病岛中从hp0541到hp0547的大片段基因组缺失。FLP - FRT重组系统在幽门螺杆菌中构建无痕缺失的创新性应用将为进一步研究假定致病基因的功能提供有用的工具，并有助于理解幽门螺杆菌的致病机制。
Helicobacter pylori is a Gram-negative, microaerophilic bacterium associated with human gastric diseases. Further investigations on virulence genes are still required to clarify the pathogenic mechanism of H. pylori and the heterogeneous problem of infection. In order to develop an efficient and accurate method to study gene functions in H. pylori pathogenesis, an unmarked deletion method for both a single gene and a large fragment was established based on the FLP-FRT recombination system. Using this method, the gene hp0788, encoding an outer membrane protein (HofF), was deleted. Deletion of hp0788 did not affect growth or motility of H. pylori, but reduced the adherence of the bacteria to gastric epithelial cells. The apoptosis of GES-1 cells caused by H. pylori infection was also reduced by the defection of hp0788. These suggest that hp0788 takes part in the bacterium-host interaction and plays an important role in H. pylori infection. Furthermore, a large genomic fragment deletion from hp0541 to hp0547 in cag pathogenicity island was also successfully achieved using FLP-FRT method. The innovative application of the FLP-FRT recombination system in H. pylori to construct unmarked deletion would provide a helpful tool for further function research of putative pathogenic genes and contribute to the understanding of H. pylori pathogenesis.
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影响因子：
4.2
作者：
Ishikawa M;Hori K
通讯作者：
Hori K
影响因子：
3.4
作者：
Backert, S;Ziska, E;Meyer, TF
通讯作者：
Meyer, TF
影响因子：
4.4
作者：
Cheng, Liu;Mirko, Rossi;Annemieke, Smet
通讯作者：
Annemieke, Smet
影响因子：
3.1
作者：
Chiang, SL;Mekalanos, JJ
通讯作者：
Mekalanos, JJ
幽门螺杆菌胃上皮细胞粘附和 CagA 注射的分子机制。
DOI：
10.1186/1478-811x-9-28
发表时间：
2011-11-01
期刊：
Cell communication and signaling : CCS
影响因子：
--
作者：
Backert S;Clyne M;Tegtmeyer N
通讯作者：
Tegtmeyer N