Bacterial artificial chromosomes as analytical basis for gene transcriptional machineries
| Autor: | Youhei W. Terakawa, Takayoshi Inoue, Yukiko U. Inoue, Junko Asami, Saki F. Egusa |
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| Rok vydání: | 2010 |
| Předmět: |
Chromosomes
Artificial Bacterial Transcription Genetic Genetic Vectors Transposon tagging Mice Transgenic Regulatory Sequences Nucleic Acid Biology Cdh6 (cadherin-6) Mice Transgenic mouse Genes Reporter Genetics Animals Humans Homologous recombination Enhancer Transposon Gene Recombination Genetic Original Paper Bacterial artificial chromosome (BAC) Bacterial artificial chromosome Genome Gene Expression Profiling Cadherins Gene expression profiling Regulatory sequence Gene transcriptional regulation DNA Transposable Elements Animal Science and Zoology Agronomy and Crop Science Functional genomics Biotechnology |
| Zdroj: | Transgenic Research |
| ISSN: | 1573-9368 0962-8819 |
| DOI: | 10.1007/s11248-010-9469-3 |
| Popis: | Bacterial Artificial Chromosomes (BACs) had been minimal components of various genome-sequencing projects, constituting perfect analytical basis for functional genomics. Here we describe an enhancer screening strategy in which BAC clones that cover any genomic segments of interest are modified to harbor a reporter cassette by transposon tagging, then processed to carry selected combinations of gene regulatory modules by homologous recombination mediated systematic deletions. Such engineered BAC-reporter constructs in bacterial cells are ready for efficient transgenesis in mice to evaluate activities of gene regulatory modules intact or absent in the constructs. By utilizing the strategy, we could speedily identify a critical genomic fragment for spatio-temporally regulated expression of a mouse cadherin gene whose structure is extraordinarily huge and intricate. This BAC-based methodology would hence provide a novel screening platform for gene transcriptional machineries that dynamically fluctuate during development, pathogenesis and/or evolution. |
| Databáze: | OpenAIRE |
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