IHB OpenIR  > 鱼类生物学及渔业生物技术研究中心  > 期刊论文
Construction of two selectable markers for integrative/conjugative plasmids in Flavobacterium columnare
Zhang Jin1,2; Zou Hong1; Wang Liangfa1; Huang Bei1; Li Nan1; Wang Guitang1; Nie Pin1; Nie, P (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2012-03-01
Source PublicationCHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY
ISSN0254-4059
Volume30Issue:2Pages:269-278
AbstractFlavobacterium columnare, the etiological agent of columnaris disease, is one of the most important and widespread bacterial pathogens of freshwater fish. In this study, we constructed two artificial selectable markers (chloramphenicol and spectinomycin resistance) for gene transfer in F. columnare. These two new artificial selectable markers, which were created by placing the chloramphenicol or spectinomycin resistance gene under the control of the native acs regulatory region of F. columnare, were functional in both F. columnare and Escherichia coli. The integrative/conjugative plasmids constructed by using these markers were introduced into F. columnare G(4) via electroporation or conjugation. The integrated plasmid DNA was confirmed by Southern blotting and PCR analysis. These two markers can be employed in future investigations into gene deletion and the pathogenicity of virulence factors in F. columnare.; Flavobacterium columnare, the etiological agent of columnaris disease, is one of the most important and widespread bacterial pathogens of freshwater fish. In this study, we constructed two artificial selectable markers (chloramphenicol and spectinomycin resistance) for gene transfer in F. columnare. These two new artificial selectable markers, which were created by placing the chloramphenicol or spectinomycin resistance gene under the control of the native acs regulatory region of F. columnare, were functional in both F. columnare and Escherichia coli. The integrative/conjugative plasmids constructed by using these markers were introduced into F. columnare G(4) via electroporation or conjugation. The integrated plasmid DNA was confirmed by Southern blotting and PCR analysis. These two markers can be employed in future investigations into gene deletion and the pathogenicity of virulence factors in F. columnare.
SubtypeArticle
KeywordFlavobacterium Columnare Selectable Marker Integrative/conjugative Plasmid Gene Disruption
Department[Zhang Jin; Zou Hong; Wang Liangfa; Huang Bei; Li Nan; Wang Guitang; Nie Pin] Chinese Acad Sci, Inst Hydrobiol, State Key Lab
DOI10.1007/s00343-012-1077-z
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
Funding OrganizationChinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University ; Chinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University ; Chinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University ; Chinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University
Indexed BySCI
Language英语
WOS Research AreaMarine & Freshwater Biology ; Oceanography
WOS SubjectLimnology ; Oceanography
WOS IDWOS:000301001900011
WOS KeywordCHONDROITIN AC LYASE ; CATFISH ICTALURUS-PUNCTATUS ; GLIDING MOTILITY ; CHANNEL CATFISH ; FLEXIBACTER-COLUMNARIS ; GENETIC MANIPULATION ; BACTEROIDES-FRAGILIS ; CYTOPHAGA JOHNSONAE ; ESCHERICHIA-COLI ; LOW VIRULENCE
Funding OrganizationChinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University ; Chinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University ; Chinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University ; Chinese Ministry of Science and Technology[2011CB944600]; US Department of Energy[DE-FG02-02ER15332]; Fudan University; Rijk Zwaan; Institute of Hydrobiology; Biology Department; Huck Institutes of the Life Sciences at the Pennsylvania State University
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Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/16741
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Corresponding AuthorNie, P (reprint author), Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
Affiliation1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Zhang Jin,Zou Hong,Wang Liangfa,et al. Construction of two selectable markers for integrative/conjugative plasmids in Flavobacterium columnare[J]. CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY,2012,30(2):269-278.
APA Zhang Jin.,Zou Hong.,Wang Liangfa.,Huang Bei.,Li Nan.,...&Nie, P .(2012).Construction of two selectable markers for integrative/conjugative plasmids in Flavobacterium columnare.CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY,30(2),269-278.
MLA Zhang Jin,et al."Construction of two selectable markers for integrative/conjugative plasmids in Flavobacterium columnare".CHINESE JOURNAL OF OCEANOLOGY AND LIMNOLOGY 30.2(2012):269-278.
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