| Characterization of eastern oyster (Crassostrea virginica Gmelin) chromosomes by fluorescence in situ hybridization with bacteriophage P1 clones | |
| Wang, YP; Xu, Z; Pierce, JC; Guo, XM | |
| 2005-05-01 | |
| 发表期刊 | MARINE BIOTECHNOLOGY
 |
| ISSN | 1436-2228 |
| 卷号 | 7期号:3页码:207-214 |
| 文章类型 | Article |
| 摘要 | 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.; 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. |
| 关键词 | Fish P1 Clones Physical Mapping Chromosome Genome Mollusca |
| 学科领域 | Biotechnology & Applied Microbiology ; Marine & Freshwater Biology |
| DOI | 10.1007/s10126-004-0051-y |
| URL | 查看原文 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000230431000008 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qdio.ac.cn/handle/337002/3536 |
| 专题 | 海洋生物技术研发中心 |
| 作者单位 | 1.Rutgers State Univ, Inst Marine & Coastal Sci, Haskins Shellfish Res Lab, Port Norris, NJ 08349 USA 2.Chinese Acad Sci, Inst Oceanol, Expt Marine Biol Lab, Qingdao 266071, Shandong, Peoples R China 3.Univ Sci Philadelphia, Dept Biol Sci, Philadelphia, PA 19104 USA |
| 推荐引用方式 GB/T 7714 | Wang, YP,Xu, Z,Pierce, JC,et al. Characterization of eastern oyster (Crassostrea virginica Gmelin) chromosomes by fluorescence in situ hybridization with bacteriophage P1 clones[J]. MARINE BIOTECHNOLOGY,2005,7(3):207-214. |
| APA | Wang, YP,Xu, Z,Pierce, JC,&Guo, XM.(2005).Characterization of eastern oyster (Crassostrea virginica Gmelin) chromosomes by fluorescence in situ hybridization with bacteriophage P1 clones.MARINE BIOTECHNOLOGY,7(3),207-214. |
| MLA | Wang, YP,et al."Characterization of eastern oyster (Crassostrea virginica Gmelin) chromosomes by fluorescence in situ hybridization with bacteriophage P1 clones".MARINE BIOTECHNOLOGY 7.3(2005):207-214. |
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