PUBLICATION
Construction and expression analysis of the zebrafish heart-specific transgenetic vector based on Tol2 transposable element
- Authors
- Chen, T., Luo, N., Xie, H., Wu, X., and Deng, Y.
- ID
- ZDB-PUB-100504-17
- Date
- 2010
- Source
- Sheng wu gong cheng xue bao = Chinese journal of biotechnology 26(2): 230-236 (Journal)
- Registered Authors
- Deng, Yun, Wu, Xiushan
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Genetically Modified/genetics
- Animals, Genetically Modified/growth & development
- DNA Transposable Elements/genetics*
- Genes, Reporter/genetics
- Genetic Vectors/biosynthesis
- Genetic Vectors/genetics*
- Green Fluorescent Proteins/genetics
- Myocardium/metabolism*
- Plasmids/genetics
- Transfection
- Transgenes
- Transposases/genetics
- Zebrafish/genetics*
- Zebrafish Proteins/genetics*
- Zebrafish Proteins/metabolism
- PubMed
- 20432943
Citation
Chen, T., Luo, N., Xie, H., Wu, X., and Deng, Y. (2010) Construction and expression analysis of the zebrafish heart-specific transgenetic vector based on Tol2 transposable element. Sheng wu gong cheng xue bao = Chinese journal of biotechnology. 26(2):230-236.
Abstract
In an effort to generate a desired expression construct for making heart-specific expression transgenic zebrafish, a Tol2 plasmid, which can drive EGFP reporter gene specifically expressed in the heart, was modified using subcloning technology. An IRES fragment bearing multiple cloning site (MCS) was amplified directly from pIRES2-EGFP plasmid and was inserted between the CMLC2 promoter and EGFP fragment of the pDestTol2CG vector. This recombinant expression plasmid pTol2-CMLC2-IRES-EGFP can drive any interested gene specifically expressed in the zebrafish heart along with EGFP reporter gene. To test the effectiveness of this new expression plasmid, we constructed pTol2-CMLC2-RED-IRES-EGFP plasmid by inserting another reporter gene DsRed-Monome into MCS downstream of the CMLC2 promoter and injected this transgenic recombinant plasmid into one-cell stage embryos of zebrafish. Under fluorescence microscope, both the red fluorescence and the green fluorescence produced by pTol2-CMLC2-RED-IRES-EGFP were detected specifically in the heart tissue in the same expression pattern. This novel expression construct pTol2-CMLC2-IRES-EGFP will become an important tool for our research on identifying heart development candidate genes' function using zebrafish as a model.
Errata / Notes
Article in Chinese.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping