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Isolation and Characterization of Male-Specific DNA Markers in the Rock Bream Oplegnathus fasciatus

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Abstract

Sex-specific DNA markers applicable were very useful for elucidating the sex-determination mechanism and sex control in fishes. In the present study, amplified fragment-length polymorphism (AFLP) approach with 144 primer combinations was employed to identify sex-specific markers in the rock bream. Four male-specific AFLP fragments were identified which were designated as Opl286, Opl237, Opl422, and Opl228. Further sequence analysis of the sex markers’ genomic region revealed subtle differences between the males and females. We identified four male-specific single-nucleotide polymorphisms (SNPs) and a deletion of 8 bp in marker Opl286, six male-specific SNPs in marker Opl237, three male-specific SNPs in marker Opl422, and eight male-specific SNPs and 1 bp inversions in marker Opl228. Specific primers were designed based on the nucleotide variation in the sequences to develop a simple polymerase chain reaction method for identifying the genetic sex of rock bream. As a result, three out of the four male-specific markers were converted into SNP markers. The male-specific AFLP markers and AFLP-derived SNP markers were tested in 100 individuals collected from three locations around the coast of Zhoushan, yielding reproducible sex identification. These male-specific DNA markers are a useful tool for the identification of the sex-determining locus in rock bream and for guiding artificial breeding programs.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (41106114), the National Science and Technology Pillar Program of China (2011BAD13B08), and the Project of Zhejiang Province of China (2010R50025; 2010F20006).

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Authors and Affiliations

  1. Key Lab of Mariculture and Enhancement of Zhejiang Province, Marine Fishery Institute of Zhejiang Province, 316100, Zhoushan, People’s Republic of China

    Dongdong Xu, Bao Lou & Hanxiang Xu

  2. Marine and Fishery Institute, Zhejiang Ocean University, 316100, Zhoushan, Zhejiang Province, People’s Republic of China

    Dongdong Xu, Sanlei Li & Zhi Geng

Authors
  1. Dongdong Xu
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  2. Bao Lou
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  3. Hanxiang Xu
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  4. Sanlei Li
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  5. Zhi Geng
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Correspondence to Dongdong Xu or Bao Lou.

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Xu, D., Lou, B., Xu, H. et al. Isolation and Characterization of Male-Specific DNA Markers in the Rock Bream Oplegnathus fasciatus . Mar Biotechnol 15, 221–229 (2013). https://doi.org/10.1007/s10126-012-9480-1

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  • DOI: https://doi.org/10.1007/s10126-012-9480-1

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