IHB OpenIR  > 鱼类生物学及渔业生物技术研究中心  > 期刊论文
Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos
Dai, Jun; Cui, Xiaojuan; Zhu, Zuoyan; Hu, Wei; Hu, W, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; huwei@ihb.ac.cn
2010
Source PublicationINTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES
ISSN1449-2288
Volume6Issue:7Pages:756-768
AbstractThis study focused on concatemer formation and integration pattern of transgenes in zebrafish embryos. A reporter plasmid based on enhanced green fluorescent protein (eGFP) driven by Cytomegalovirus (CMV) promoter, pCMV-pax6in-eGFP, was constructed to reflect transgene behavior in the host environment. After removal of the insertion fragment by double digestion with various combinations of restriction enzymes, linearized pCMV-pax6in-eGFP vectors were generated with different combinations of 5'-protruding, 3'-protruding, and blunt ends that were microinjected into zebrafish embryos. Repair of double-strand breaks (DSBs) was monitored by GFP expression following religation of the reporter gene. One-hundred-and-ninety-seven DNA fragments were amplified from GFP-positive embryos and sequenced to analyze the repair characteristics of different DSB end combinations. DSBs involving blunt and asymmetric protruding ends were repaired efficiently by direct ligation of blunt ends, ligation after blunting and fill-in, or re moved by cutting. Repair of DSBs with symmetric 3'-3' protrusions was less efficient and utilized template-directed repair. The results suggest that non-homologous end joining (NHEJ) was the principal mechanism of exogenous gene concatemer formation and integration of transgenes into the genome of transgenic zebrafish.
KeywordTransgene Concatemer Dsb (Double Strand Breaks) Nhej (Non-homologous End Joining) Zebrafish
Department[Dai, Jun; Cui, Xiaojuan; Zhu, Zuoyan; Hu, Wei] Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; [Dai, Jun; Cui, Xiaojuan] Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
Subject AreaBiochemistry & Molecular Biology ; Biology
Funding Organizationstate Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141] ; state Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141] ; state Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141] ; state Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141]
Indexed BySCI
Language英语
WOS IDWOS:000285770600014
Funding Organizationstate Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141] ; state Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141] ; state Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141] ; state Key Fundamental Research of China [2007CB109206]; National Natural Science Foundation [30930069, 30430540]; '863' High Technology Project of China [2006AA10Z141]
Citation statistics
Cited Times:27[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/15494
Collection鱼类生物学及渔业生物技术研究中心_期刊论文
Corresponding AuthorHu, W, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; huwei@ihb.ac.cn
Recommended Citation
GB/T 7714
Dai, Jun,Cui, Xiaojuan,Zhu, Zuoyan,et al. Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos[J]. INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES,2010,6(7):756-768.
APA Dai, Jun,Cui, Xiaojuan,Zhu, Zuoyan,Hu, Wei,Hu, W, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China,&huwei@ihb.ac.cn.(2010).Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos.INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES,6(7),756-768.
MLA Dai, Jun,et al."Non-Homologous End Joining Plays a Key Role in Transgene Concatemer Formation in Transgenic Zebrafish Embryos".INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES 6.7(2010):756-768.
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