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Title: LINKAGE MAPPING AND COMPARATIVE ANALYSIS OF BOVINE EXPRESSED SEQUENCE TAGS (ESTS)

Author
item Grosse, William
item Kappes, Steven - Steve
item MCGRAW, R - UNIVERSITY OF GEORGIA

Submitted to: Animal Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/22/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Bovine expressed sequence tags (ESTs) containing microsatellites are potentially suitable markers for both linkage and comparative maps. These gene markers can be used to link the physical and linkage maps and provide a way to compare gene locations across different species. Given the vast amount of human gene mapping and gene trait information, such as disease genes, economically important candidate genes can be identified in cattle based on their gene function in humans. We isolated cDNA clones, representing expressed gene, using a microsatellite (CA)10 probe. The genes were used to search the DNA databases in GenBank to identify orthologs to the bovine genes. Predicted bovine map positions, where possible, were obtained using human/bovine comparative mapping data. Isolated bovine microsatelites were used to place these genes on the bovine linkage map. The results demonstrated that mapping bovine ESTs containing microsatellites is a plausible strategy to increase the density of gene markers on the bovine linkage and comparative maps.

Technical Abstract: Bovine expressed sequence tags (ESTs) containing microsatellites are suitable markers for both linkage and comparative maps. We isolated clones from a bovine fetal thigh skeletal muscle cDNA library that were positive for a (CA)10 probe. Thirty individual clones were isolated and characterized by sequencing. Sequences from the 5' and 3' ends of a clone were considered as separate ESTs until a contiguous sequence was identified. A total of 47 ESTs were sequenced from the 5' and/or 3' ends and full sequence was obtained for the 30 clones. BLASTn analysis identified significant homology to known mammalian coding regions for 31 of the bovine ESTs, 30 of which also matched human ESTs or sequence tag sights (STS). The remaining 16 bovine ESTs represented novel transcripts. Microsatellites were isolated in 27 of the ESTs, 11 of which were developed into markers and placed on the MARC bovine linkage map. Human cytogenetic map positions were available for 20 of the 30 human EST orthologs, and a putative bovine map position for 17 of the sequences could be inferred using comparative mapping data. These results demonstrated that mapping bovine ESTs containing microsatellites is a plausible strategy to increase the density of gene markers on the bovine linkage and comparative maps.