Physically mapped, cosmid-derived microsatellite markers as anchor loci on bovine chromosomes

Abstract

To identify physical and genetic anchor loci on bovine chromosomes, 13 cosmids, obtained after the screening of partial bovine cosmid libraries with the (CA)n microsatellite motif, were mapped by fluorescence in situ hybridization (FISH). Eleven cosmid probes yielded a specific signal on one of the bovine chromosomes and identified the following loci: D5S2, D5S3, D6S3, D8S1, D11S5, D13S1, D16S5, D17S2, D19S2, D19S3, D21S8. Two cosmids produced centromeric signals on many chromosomes. The microsatellite-containing regions were subcloned and sequenced. The sequence information revealed that the two centromeric cosmids were derived from bovine satellites 1.723 and 1.709, respectively. A cosmid located in the subtelomeric region of Chromosome (Chr) 17 (D17S2) had features of a chromosome-specific satellite. Primers were designed for eight of the nonsatellite cosmids, and seven of these microsatellites were polymorphic with between three and eight alleles on a set of outbred reference families. The polymorphic and chromosomally mapped loci can now be used to physically anchor other bovine polymorphic markers by linkage analysis. The microsatellite primers were also applied to DNA samples of a previously characterized panel of somatic hybrid cell lines, allowing the assignment of seven microsatellite loci to defined syntenic groups. These assignments confirmed earlier mapping results, revealed a probable case of false synteny, and placed two formerly unassigned syntenic groups on specific chromosomes.

Low diversity in the major histocompatibility complex class II DRB1 gene of the Spanish ibex, Capra pyrenaica

“…These were : INRA005, INRA023, INRA032, INRA035, INRA037, INRA063, CHIAE54, CSSM66, HEL9, ILST005, ETH10, ETH152, ETH225. Primer identification, PCR conditions and amplification profiles for each microsatellite locus are described elsewhere (Brezinsky et al, 1993;Kaukinen and Varvio, 1993;Steffen et al, 1993;Toldo et al, 1993;Barendse et al, 1994;Vaiman et al, 1994;Amills et al, 1996b). Fluorescent genotyping was performed in an ABI Prism 310 automated DNA sequencing system (Perkin Elmer Biosystems, Foster City, CA 94404, USA).…”

Genome mapping in ruminants and map locations for genes influencing reproduction

“…The fluorescent tag is bound to the probe and detected with a sensitive ultraviolet microscope. Large cloned DNA fragments (cosmids) containing inhibin genes were used in FISH experiments to assign the βA-inhibin gene to cattle chromosome 4 (Neibergs et al, 1993) and the βB-inhibin gene to sheep and cattle chromosome 2 (Goldammer et al, 1995).…”

“…At least one marker from each group must be mapped by physical assignment to determine the chromosome corresponding to each linkage group. FISH analysis, using DNA sequences that contain framework markers from the linkage map, has been used to anchor the linkage and physical maps of bovine chromosomes (Hawkins et al, 1995;Toldo et al, 1993;Ferretti et al, 1997).…”

Allele frequencies of microsatellite loci for genetic characterization of a Sicilian bovine population

“…In 2006, the International Society for Animal Genetics (ISAG) recommended a panel of nine microsatellite loci for cattle genetic analysis: TGLA227 (Georges and Massey, 1992), BM2113 (Sunden et al, 1993), ETH10 (Toldo et al, 1993), SPS115 (Baylor College of Medicine Human Genome Sequencing Center, 2006), TGLA126 (Georges and Massey, 1992), TGLA122 (Georges and Massey, 1992), INRA023 (Vaiman et al, 1994), ETH225 , and BM1824 (Barendse et al, 1994). In 2008, the ISAG added three additional microsatellite loci: BM1818 (Bishop et al, 1994), ETH3 (Toldo et al, 1993), and TGLA53 (Georges and Massey, 1992). The aim of this study was to test the 11 microsatellite markers that were suggested by the ISAG and to evaluate genetic variability to create a database of allele frequencies and improve parental definitions and genetic traceability of the Sicilian cattle population.…”