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Characterization of Eastern Oyster (Crassostrea virginica Gmelin) Chromosomes by Fluorescence In Situ Hybridization with Bacteriophage P1 Clones

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Abstract

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.

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Acknowledgments

This study was partly supported by grants from National Sea Grant (B/T-9801, R/OD-2003-1), U.S. Department of Agriculture NRI (96-35205-3854), and the New Jersey Commission on Science and Technology (02-2042-007-11). Y.W. was a visiting student from Institute of Oceanology, Chinese Academy of Sciences, and partly supported by grants from National Science Foundation of China (39825121), the 863 Program (2001AA628150), and the 973 Program (G1999012007). This is contribution IMCS-2004-20 and NJMSC-04-570.

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Authors and Affiliations

  1. Haskin Shellfish Research Laboratory, Institute of Marine and Coastal Sciences, Rutgers University, 6959 Miller Avenue, Port Norris, NJ, 08349, U.S.A.

    Yongping Wang, Zhe Xu & Ximing Guo

  2. Experimental Marine Biology Laboratory, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, Shandong, 266071, P.R.C.

    Yongping Wang

  3. Department of Biological Sciences, University of the Sciences in Philadelphia, Philadelphia, PA, 19104, U.S.A.

    James C. Pierce

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  1. Yongping Wang
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  4. Ximing Guo
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Correspondence to Ximing Guo.

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Wang, Y., Xu, Z., Pierce, J.C. et al. Characterization of Eastern Oyster (Crassostrea virginica Gmelin) Chromosomes by Fluorescence In Situ Hybridization with Bacteriophage P1 Clones. Mar Biotechnol 7, 207–214 (2005). https://doi.org/10.1007/s10126-004-0051-y

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