Targeted development of microsatellite markers from the defined region of bovine chromosome 6q21-31.
A methodical strategy for the isolation of microsatellite markers specific for targeted regions of bovine chromosomes is presented. The procedure involves directed microdissection of one defined subchromosomal area, its DOP-PCR-amplification and cloning. With this approach, a library specific to the BTA 6q21-31 chromosomal region was constructed. Eleven unique microsatellite-containing sequences were isolated, converted into sequence-tagged microsatellite sites, and characterized concerning their species-specific origin. Seven primer pairs generated bovine-specific PCR products and provided a set of microsatellite markers that generally revealed high informativity in the HF breed. Linkage analysis assigned six of them to their predefined subchromosomal origin on BTA 6 corresponding to the specific rehybridization signal of the DOP-PCR product generated from the microdissected chromosome area 6q21-31. The results underline the usefulness of the BTA 6q21-31 library for targeted isolation of unique sequences that are specific for the dissected chromosomal region as demonstrated here by the isolation of microsatellite markers.
Weikard
R
R
Forschungsinstitut für die Biologie landwirtschaftlicher Nutztiere, Forschungsbereich Molekularbiologie, Dummerstorf, Germany.
Goldammer
T
T
Kühn
C
C
Barendse
W
W
Schwerin
M
M
D000818:Animals; D002417:Cattle / Q000235:genetics*; D002874:Chromosome Mapping / Q000662:veterinary*; D005260:Female; D015723:Gene Library; D007178:Inbreeding; D008297:Male; D018895:Microsatellite Repeats; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic