BAC constructs in transgenic reporter mouse lines control efficient and specific LacZ expression in hypertrophic chondrocytes under the complete Col10a1 promoter
| Autor: | Nadia Adam, Klaus von der Mark, Eva Bauer, Britta Schlund, Benoit de Crombrugghe, Ernst Pöschl, Takako Hattori, Sonja Gebhard, Michael R. Bösl |
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| Rok vydání: | 2006 |
| Předmět: |
Genetically modified mouse
Chromosomes Artificial Bacterial Histology Genotype Transgene Gene Expression Mice Transgenic Biology Mice 03 medical and health sciences Exon Chondrocytes Genes Reporter Osteogenesis Transgenic mouse Gene expression Animals Growth Plate Transgenes Homologous recombination Promoter Regions Genetic Molecular Biology Gene Endochondral ossification BAC 030304 developmental biology Recombination Genetic Original Paper 0303 health sciences Reporter gene Bacterial artificial chromosome 030302 biochemistry & molecular biology Gene Transfer Techniques Cell Biology Embryo Mammalian Molecular biology Medical Laboratory Technology Lac Operon Hypertrophic cartilage Collagen X Collagen Type X |
| Zdroj: | Histochemistry and Cell Biology |
| ISSN: | 1432-119X 0948-6143 |
| DOI: | 10.1007/s00418-006-0236-8 |
| Popis: | During endochondral ossification hypertrophic chondrocytes in the growth plate of fetal long bones, ribs and vertebrae play a key role in preparing growth plate cartilage for replacement by bone. In order to establish a reporter gene mouse to facilitate functional analysis of genes expressed in hypertrophic chondrocytes in this process, Col10a1- BAC reporter gene mouse lines were established expressing LacZ specifically in hypertrophic cartilage under the control of the complete Col10a1 gene. For this purpose, a bacterial artificial chromosome (BAC RP23-192A7) containing the entire murine Col10a1 gene together with 200 kb flanking sequences was modified by inserting a LacZ-Neo cassette into the second exon of Col10a1 by homologous recombination in E. coli. Transgenic mice containing between one and seven transgene copies were generated by injection of the purified BAC-Col10a1- lLacZ DNA. X-gal staining of newborns and embryos revealed strong and robust LacZ activity exclusively in hypertrophic cartilage of the fetal and neonatal skeleton of the transgenic offspring. This indicates that expression of the reporter gene in its proper genomic context in the BAC Col10a1 environment is independent of the integration site and reflects authentic Col10a1 expression in vivo. The Col10a1 specific BAC recombination vector described here will enable the specific analysis of effector gene functions in hypertrophic cartilage during skeletal development, endochondral ossification, and fracture callus healing. |
| Databáze: | OpenAIRE |
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