A recombinogenic targeting method to modify large-inserts for cis -regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene
| Autor: | Janet L. Carr, Frank H. Ruddle, Joseph K. Hwang, Cooduvalli S. Shashikant, Jaya Bhargava, Chi-hua Chiu, Günter P. Wagner, Chris T. Amemiya |
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| Rok vydání: | 2000 |
| Předmět: |
Homeodomain Proteins
Regulation of gene expression Reporter gene biology Transgene Genetic Vectors DNA Recombinant Gene Expression Regulation Developmental Mice Transgenic Zebrafish Proteins biology.organism_classification Molecular biology Mice Exon Lac Operon Shuttle vector Genes Reporter Gene Targeting Gene expression Genetics Animals Gene Zebrafish Developmental Biology |
| Zdroj: | Development Genes and Evolution. 210:105-109 |
| ISSN: | 1432-041X 0949-944X |
| DOI: | 10.1007/s004270050016 |
| Popis: | The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing a portion of the zebrafish HoxA-b cluster, was captured into the yeast-bacterial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ reporter gene was then inserted in-frame into exon 1 of the zfHoxa-11b locus by a second round of recombinogenic targeting. Expression of the zfHoxa-11b-lacZ reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only in the posterior portion of the A-P axis, in the paraxial mesoderm, neural tube, and somites. These findings demonstrate the utility of recombinogenic targeting for the modification and expression of large inserts captured from P1/PAC clones. |
| Databáze: | OpenAIRE |
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