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

  • Xiaojun Zhang
  • Yang Zhang
  • Chantel Scheuring
  • Hong-Bin Zhang
  • Pin Huan
  • Bing Wang
  • Chengzhang Liu
  • Fuhua Li
  • Bin Liu
  • Jianhai Xiang
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

Authors and Affiliations

  • Xiaojun Zhang
    • 1
  • Yang Zhang
    • 1
    • 2
    • 3
  • Chantel Scheuring
    • 3
  • Hong-Bin Zhang
    • 3
  • Pin Huan
    • 1
    • 2
  • Bing Wang
    • 1
  • Chengzhang Liu
    • 1
  • Fuhua Li
    • 1
  • Bin Liu
    • 1
  • Jianhai Xiang
    • 1
  1. 1.Institute of OceanologyChinese Academy of SciencesQingdaoChina
  2. 2.Graduate University of the Chinese Academy of SciencesBeijingChina
  3. 3.Department of Soil & Crop SciencesTexas A&M UniversityCollege StationUSA