Targeted development of microsatellite markers from the defined region of bovine Chromosome 6q21-31 |
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Authors: | Rosemarie Weikard T Goldammer Ch Kühn W Barendse M Schwerin |
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Institution: | (1) Forschungsinstitut für die Biologie landwirtschaftlicher Nutztiere, Forschungsbereich Molekularbiologie, Wilhelm-Stahl-Allee 2, D-18196 Dummerstorf, Germany, DE;(2) CSIRO Tropical Agriculture, Gehrman Labs, University of Queensland, St. Lucia 4072, Australia, AU |
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Abstract: | 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.
Received: 27 March 1997 / Accepted: 14 July 1997 |
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