Plant & Animal Genome VI Conference

P69

ISOLATION OF HIGHLY POLYMORPHIC MICROSATELLITES IN ALPACA FOR GENOMIC CHARACTERIZATION OF SOUTH-AMERICAN CAMELIDS.

1. Lab. Biotechnology, INIA La Platina, P.O.Box 439/3, Santiago, CHILE
2. Dept. of Veterinaty Science, University of Kentucky, Lexington, KY 40546, USA
3. M.H. Gluck Equine Research Center, Univ. of Kentucky, Lexington KY 40546

Simple Sequence Repeats (SSRs) or microsatellites exhibit the highest polymorphic index among the most commonly used markers for genomic analysis. This high variability comes from the polymorphism detected in the number of repeats of the di-, tri- or tetra-nucleotides at each SSR locus. With the aim of identifying new SSRs in South American Camelids (CSA), a genomic library was prepared on the pLITMUS cloning vector, selecting fragments between 400-bp and 700-bp. Almost 100 clones of this library showed positive hybridyzation against a (dA-dC)_n(dG-dT)_n probe. The complete sequencing of these clones elucidated 57 clones having (TG) repetitions, with variation in length and complexity. Using the "Oligo v. 5.0" program, primer sets were designed for 23 of these SSR clones. Of this group, five primer sets have rendered appropriate PCR products. The preliminary testing of these markers on a group of 35 alpacas have showed that these SSRs were highly polymorphic, with a large number of alleles. Additional primer sets are now under evaluation. These markers will be used to examine the extent of genetic differentiation of the four CSA species and also will be used in the development of a genetic map of these species. Financed by Fundación FIA-Chile (Project #016-94).