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Upstream regulatory region of zebrafish lunatic fringe: Isolation and promoter analysis
Liu, Jing; Sun, Yong-Hua; Wang, Na; Wang, Ya-Ping; Zhu, Zuo-Yan; Zhu, ZY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
2006-07-01
Source PublicationMARINE BIOTECHNOLOGY
ISSN1436-2228
Volume8Issue:4Pages:357-365
AbstractLunatic fringe (Lfng), one modulator of Notch signaling, plays an essential part in demarcation of tissues boundaries during animal early development, especially somitogenesis. To characterize the promoter of zebrafish 1fng and generate somite-specific transgenic zebrafish, we isolated the upstream regulatory region of zebrafish 1fng by blast search at the Ensembl genome database (http://www. ensembl.org) and analyzed the promoter activity using green fluorescent protein (GFP) as a reporter. Promoter activity assay in zebrafish shows that the 0.2-kb fragment containing GC-box, CAAT-box, and TATA-box can direct tissue-specific GFP expression, while the 0.4-kb and 1.2-kb fragments with further upstream sequence included drive GFP expression more efficiently. We produced 1fngEGFP-transgenic founders showing somite-specific expression of GFP and consequently generated a hemizygous 1fngEGFP-transgenic line. The eggs from 1fngEGFP-transgenic female zebrafish show strong GFP expression, which is consistent to the reverse-transcription polymerase chain reaction PCR (RT-PCR) detection of 1fng transcripts in the fertilized eggs. This reveals that zebrafish 1fng is a maternal factor existing in matured eggs, suggesting that fish somitogcnesis may be influenced by maternal factors.; Lunatic fringe (Lfng), one modulator of Notch signaling, plays an essential part in demarcation of tissues boundaries during animal early development, especially somitogenesis. To characterize the promoter of zebrafish 1fng and generate somite-specific transgenic zebrafish, we isolated the upstream regulatory region of zebrafish 1fng by blast search at the Ensembl genome database (http://www. ensembl.org) and analyzed the promoter activity using green fluorescent protein (GFP) as a reporter. Promoter activity assay in zebrafish shows that the 0.2-kb fragment containing GC-box, CAAT-box, and TATA-box can direct tissue-specific GFP expression, while the 0.4-kb and 1.2-kb fragments with further upstream sequence included drive GFP expression more efficiently. We produced 1fngEGFP-transgenic founders showing somite-specific expression of GFP and consequently generated a hemizygous 1fngEGFP-transgenic line. The eggs from 1fngEGFP-transgenic female zebrafish show strong GFP expression, which is consistent to the reverse-transcription polymerase chain reaction PCR (RT-PCR) detection of 1fng transcripts in the fertilized eggs. This reveals that zebrafish 1fng is a maternal factor existing in matured eggs, suggesting that fish somitogcnesis may be influenced by maternal factors.
SubtypeArticle
KeywordGreen Fluorescent Protein Lfng Maternal Factor Promoter Zebrafish
DepartmentChinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China; Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
Subject AreaBiotechnology & Applied Microbiology ; Marine & Freshwater Biology
DOI10.1007/s10126-005-5125-y
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology ; Marine & Freshwater Biology
WOS SubjectBiotechnology & Applied Microbiology ; Marine & Freshwater Biology
WOS IDWOS:000239656600004
WOS KeywordGFP REPORTER GENE ; SOMITE SEGMENTATION ; TRANSGENIC ZEBRAFISH ; SIGNALING MOLECULE ; DYNAMIC EXPRESSION ; NERVOUS-SYSTEM ; RADICAL FRINGE ; NOTCH ; SOMITOGENESIS ; DORSAL
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Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/152342/8878
Collection期刊论文
Corresponding AuthorZhu, ZY, 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 Sch, Beijing 100039, Peoples R China
Recommended Citation
GB/T 7714
Liu, Jing,Sun, Yong-Hua,Wang, Na,et al. Upstream regulatory region of zebrafish lunatic fringe: Isolation and promoter analysis[J]. MARINE BIOTECHNOLOGY,2006,8(4):357-365.
APA Liu, Jing,Sun, Yong-Hua,Wang, Na,Wang, Ya-Ping,Zhu, Zuo-Yan,&Zhu, ZY, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China.(2006).Upstream regulatory region of zebrafish lunatic fringe: Isolation and promoter analysis.MARINE BIOTECHNOLOGY,8(4),357-365.
MLA Liu, Jing,et al."Upstream regulatory region of zebrafish lunatic fringe: Isolation and promoter analysis".MARINE BIOTECHNOLOGY 8.4(2006):357-365.
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