Plant & Animal Genome IX Conference

Poster: Sequencing & EST
P01_01.html

ISOLATION OF EXPRESSED SEQUENCE TAGS FROM A THOROUGHBRED HORSE (Equus caballus) 5'RACE cDNA LIBRARY

¹ Departments of Environmental and Population Health and Biological Sciences, Tufts University School of Veterinary Medicine, North Grafton, MA, 01536, USA

² Laboratorio de Citogenetica y Genetica Molecular, Facultad de Veterinaria, Universidad de Zaragoza, Miguel Servet 177, 50013 Zaragoza, Spain

Three hundred and twenty cDNAs were isolated from a cDNA library originated from blood of a Thoroughbred septic foal. Single passage sequencing using M13 reverse primer provided 181 unique sequences. The remaining clones were either redundant, with very small insert (<50 bp), unreadable, or contained no insert. Out of the 181 unique sequences, 67 showed similarities to known genes, 56 were similar to unknown genes, and 58 were novel sequences. Seven of the 67 known genes were of mitochondrial-DNA origin (ATPase, cytochrome b, NADH1-3, COI, 16s rRNA) and 60 were of nuclear-DNA origin with different functions, including those involved in cell division (histone, cyclin I, cyclin dependent kinase2), cell signalling/cell communication (IL1-ra, G-protein receptors 1&2, immunoglobulin receptor, GCSF receptor, glia-derived neurite promoting factor, c-fms protooncogene, complement C5a receptor, IGF-II receptor, Burkitt's lymphoma receptor1, EGF-like EMR-2, cell adhesion GMP140, myeloid-associated differentiation protein, protein kinase C-delta13, cleft lip and palate transmembrane protein, myocardial vascular inhibitor factor), cell structure/motility (tropomyosin, lysosome-associated membrane protein), gene/protein expression (RNA polymeraseII, Not 56-like, RING zinc finger protein, polypirimidine tract-binding protein, ribosomal L10&L18, 45S pre rRNA), metabolism (vacuolar type H+ATPase, 75Kd iron sulphur protein, CAB1, NAD+-specific isocitrate dehydrogenase, ornithine decarboxylase antyzime, lysosomal pepstatin insensitive protease), and cell/organism defense (beta lactoglobulin II, transferrin, P2XM, zinedin, y-L aminoacid transporter1, MHC class II DQ-alpha, Ig lambda, LY6H protein, HIV-EP2/Shnurri2), among others. Ninety four out of 181 clones contained microsatellites of 3 or more repeats, 98% of which had flanking sequences suitable for allele amplification by PCR. These ESTs will be a valuable addition to the horse EST database and may be useful for gene mapping and other genetic analysis in horses.