Marine Biotechnology

, Volume 12, Issue 2, pp 141–149

Construction and Characterization of a Bacterial Artificial Chromosome (BAC) Library of Pacific White Shrimp, Litopenaeus vannamei

Authors

  • Xiaojun Zhang
    • Institute of OceanologyChinese Academy of Sciences
  • Yang Zhang
    • Institute of OceanologyChinese Academy of Sciences
    • Graduate University of the Chinese Academy of Sciences
    • Department of Soil & Crop SciencesTexas A&M University
  • Chantel Scheuring
    • Department of Soil & Crop SciencesTexas A&M University
  • Hong-Bin Zhang
    • Department of Soil & Crop SciencesTexas A&M University
  • Pin Huan
    • Institute of OceanologyChinese Academy of Sciences
    • Graduate University of the Chinese Academy of Sciences
  • Bing Wang
    • Institute of OceanologyChinese Academy of Sciences
  • Chengzhang Liu
    • Institute of OceanologyChinese Academy of Sciences
  • Fuhua Li
    • Institute of OceanologyChinese Academy of Sciences
  • Bin Liu
    • Institute of OceanologyChinese Academy of Sciences
    • Institute of OceanologyChinese Academy of Sciences
Original Article

DOI: 10.1007/s10126-009-9209-y

Cite this article as:
Zhang, X., Zhang, Y., Scheuring, C. et al. Mar Biotechnol (2010) 12: 141. doi:10.1007/s10126-009-9209-y

Abstract

The pacific white shrimp (Litopenaeus vannamei) is one of the most economically important marine aquaculture species in the world. To facilitate gene cloning and characterization, genome analysis, physical mapping, and molecular selection breeding of marine shrimp, we have developed the techniques to isolate high-quality megabase-sized DNA from hemocyte nuclear DNA of female shrimp and constructed a bacterial artificial chromosome (BAC) genomic library for the species. The library was constructed in the Hind III site of the vector pECBAC1, consisting of 101,760 clones arrayed in 265 384-well microtiter plates, with an average insert size of 101 kb, and covering the genome approximately fivefold. To characterize the library, 92,160 clones were spotted onto high-density nylon filters for hybridization screening. A set of 18 pairs of overgo probes designed from eight cDNA sequences of L. vannamei genes were used in hybridization screening, and 35 positive clones were identified. These results suggest that the shrimp BAC libraries will provide a useful resource for screening of genomic regions of interest candidate genes, gene families, or large-sized synthetic DNA region and promote future works on comparative genomics, physical mapping, and large-scale genome sequencing in the species.

Keywords

Pacific white shrimpLitopenaeus vannameiBacterial artificial chromosome (BAC) libraryGenomeOvergo probe

Copyright information

© Springer Science+Business Media, LLC 2009