Preliminary Study of BAC Library Construction in Black Tiger Shrimp, Penaeus monodon

Suwit WUTHISUTHIMETHAVEE; Takashi AOKI; Ikuo HIRONO; Anchalee TASSANAKAJON

Authors

Suwit WUTHISUTHIMETHAVEE School of Agricultural Technology, Walailak University, Nakhon Si Thammarat 80161
Takashi AOKI Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology
Ikuo HIRONO Laboratory of Genome Science, Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology
Anchalee TASSANAKAJON Laboratory of Molecular Genetics and Shrimp Genomics, Faculty of Science, Chulalongkorn University, Bangkok 10330

Keywords:

BAC, Penaeus monodon, genome

Abstract

Availability of shrimp genome information is necessary for shrimp genetic studies and large-insert DNA clones, bacterial artificial chromosome (BACs) serve as valuable tools for obtaining genomic sequences. The construction of a BAC library was achieved from this preliminary study of P. monodon. High molecular weight (HMW) genomic DNA was isolated from abdominal muscle and the resulting hemocytes were of high quality and sufficient quantity for a BAC library construction. This BAC library was constructed from hemocyte HMW genomic DNA fragments ranging between 100 and 300 kb ligated to a pBAC-lac vector. The average insert size of the BAC clones was calculated as 100 kb ranging from 40 - 150 kb with over 50 % of the clones containing inserts larger than 100 kb. This BAC library contained approximately 60,000 BAC clones representing a 3-fold coverage of the haploid shrimp genome size. Furthermore, the consistency of BAC clones was also proven as no apparent rearrangements were observed before and after 100 generations in serial growth. From the above result, it is essential to improve the insert size and clone numbers of the BAC library to be a powerful tool in the future.

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