Characterization of Eastern Oyster (Crassostrea virginica Gmelin) Chromosomes by Fluorescence In Situ Hybridization with Bacteriophage P1 Clones
| Autor: | Ximing Guo, Zhe Xu, James C. Pierce, Yongping Wang |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
Applied Microbiology and Biotechnology
Genome Chromosomes chemistry.chemical_compound Gene mapping medicine Animals Nick translation Bacteriophage P1 Cloning Molecular In Situ Hybridization Fluorescence Sequence Tagged Sites Genetics biology medicine.diagnostic_test Chromosome Mapping Computational Biology Chromosome Sequence Analysis DNA biology.organism_classification Ostreidae Molecular biology chemistry Karyotyping Crassostrea Deoxyuracil Nucleotides Eastern oyster Digoxigenin DNA Fluorescence in situ hybridization |
| Zdroj: | Marine Biotechnology. 7:207-214 |
| ISSN: | 1436-2236 1436-2228 |
| DOI: | 10.1007/s10126-004-0051-y |
| Popis: | Chromosome identification is an essential step in genomic research, which so far has not been possible in oysters. We tested bacteriophage P1 clones for chromosomal identification in the eastern oyster Crassostrea virginica, using fluorescence in situ hybridization (FISH). P1 clones were labeled with digoxigenin-11-dUTP using nick translation. Hybridization was detected with fluorescein-isothiocyanate-labeled anti-digoxigenin antibodies and amplified with 2 layers of antibodies. Nine of the 21 P1 clones tested produced clear and consistent FISH signals when Cot-1 DNA was used as a blocking agent against repetitive sequences. Karyotypic analysis and cohybridization positively assigned the 9 P1 clones to 7 chromosomes. The remaining 3 chromosomes can be separated by size and arm ratio. Five of the 9 P1 clones were sequenced at both ends, providing sequence-tagged sites that can be used to integrate linkage and cytogenetic maps. One sequence is part of the bone morphogenetic protein type 1b receptor, a member of the transforming growth factor superfamily, and mapped to the telomeric region of the long arm of chromosome 2. This study shows that large-insert clones such as P1 are useful as chromosome-specific FISH probes and for gene mapping in oysters. |
| Databáze: | OpenAIRE |
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