Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis
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 acc...
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Frontiers Media S.A.
2017-11-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | http://journal.frontiersin.org/article/10.3389/fmicb.2017.02357/full |
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| author | Xiaofei Ji Ying Wang Jiaojiao Li Qianyu Rong Xingxing Chen Ying Zhang Xiaoning Liu Boqing Li Huilin Zhao |
| author_facet | Xiaofei Ji Ying Wang Jiaojiao Li Qianyu Rong Xingxing Chen Ying Zhang Xiaoning Liu Boqing Li Huilin Zhao |
| author_sort | Xiaofei Ji |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-17T01:08:22Z |
| format | Article |
| id | doaj.art-e4da6371bc58486f83120dbe222ee63e |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-17T01:08:22Z |
| publishDate | 2017-11-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-e4da6371bc58486f83120dbe222ee63e2022-12-21T22:09:13ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2017-11-01810.3389/fmicb.2017.02357263607Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in PathogenesisXiaofei Ji0Ying Wang1Jiaojiao Li2Qianyu Rong3Xingxing Chen4Ying Zhang5Xiaoning Liu6Boqing Li7Huilin Zhao8Department of Pathogenic Biology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, ChinaCentral Laboratory, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, ChinaDepartment of Pathogenic Biology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, ChinaDepartment of Pathogenic Biology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, ChinaDepartment of Pathogenic Biology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, ChinaDepartment of Pathogenic Biology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, ChinaCentral Laboratory, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, ChinaDepartment of Pathogenic Biology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, ChinaDepartment of Pathogenic Biology, School of Basic Medical Sciences, Binzhou Medical University, Yantai, ChinaHelicobacter 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.http://journal.frontiersin.org/article/10.3389/fmicb.2017.02357/fullHelicobacter pyloriunmarked deletionFLP-FRThp0788cell apoptosis |
| spellingShingle | Xiaofei Ji Ying Wang Jiaojiao Li Qianyu Rong Xingxing Chen Ying Zhang Xiaoning Liu Boqing Li Huilin Zhao Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis Frontiers in Microbiology Helicobacter pylori unmarked deletion FLP-FRT hp0788 cell apoptosis |
| title | Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis |
| title_full | Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis |
| title_fullStr | Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis |
| title_full_unstemmed | Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis |
| title_short | Application of FLP-FRT System to Construct Unmarked Deletion in Helicobacter pylori and Functional Study of Gene hp0788 in Pathogenesis |
| title_sort | application of flp frt system to construct unmarked deletion in helicobacter pylori and functional study of gene hp0788 in pathogenesis |
| topic | Helicobacter pylori unmarked deletion FLP-FRT hp0788 cell apoptosis |
| url | http://journal.frontiersin.org/article/10.3389/fmicb.2017.02357/full |
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