PAG-V: P324 - GENERATION OF EXPRESSED SEQUENCE TAGS (ESTs) FROM MUSCLE cDNA LIBRARY OF SHRIMP (<i>Penaeus vannamei</i>)

Plant & Animal Genome V Conference

Town & Country Hotel, San Diego, CA, January 12-16, 1997.

P324

GENERATION OF EXPRESSED SEQUENCE TAGS (ESTs) FROM MUSCLE cDNA LIBRARY OF SHRIMP (Penaeus vannamei)

DHAR, ARUN K., Acacia Alcivar-Warren
Department of Comparative Medicine, Tufts University School of Veterinary Medicine, 200 Westboro Road, North Grafton, MA 01536, USA

To generate expressed sequenced tags (ESTs) for identifying Type I markers in shrimp, poly(A)⁺ RNA was isolated from tail muscle of shrimp Population 2 developed by the US Marine Shrimp Farming Program Consortium. A cDNA library was constructed in lambda ZAP II vector using ZAP cDNA Gigapack III Gold cloning kit (stratagene). Mass excision of phage library was performed using ExAssist helper phage and Escherichia coli SOLAR strain. Eleven cDNA clones isoloated so far ranged in size from ~0.3 to ~6.0 kilobases. Ten of these clones were partially sequenced, with length ranging from ~152 to 231 base pairs (bp). Similarity searches for each of the partially sequenced clones were performed using FASTA program in the GenBank database. The similarity with different known genes varied from 52% to 65% with length overlaps of 49 bp to 160 bp. These included genes like Caenorhabditis elegans myosin regulation gene, C. elegans G-protein coupled receptor, C. elegans tubulin gamma chain, human fragile X mental retardation protein, human cytomegalo virus, mouse platelet derived growth factor, Clostridium perfingens hyaluronidase gene, and Saccharomyces cerevisiae DNA for ORF from chromosome XV. Two clones showed similarity to C. elegans cosmids. In summary, shrimp tail muscle cDNA library is a useful source to isolate ESTs and to identify unknkown genes. We plan to use these EST clones as probes to develop markers for mapping economically important traits like growth performance and disease resistance in cultured shrimp.