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Animal Breeding and Genetics
Asian-Australasian Journal of Animal Sciences 2007;20(3): 316-327.
DOI: https://doi.org/10.5713/ajas.2007.316    Published online January 24, 2007.
Chromosomal Localization of Korean Cattle (Hanwoo) BAC Clones via BAC end Sequence Analysis
Sung-Hwa Chae, Jae-Woo Kim, Jae Min Choi, Denis M. Larkin, Annelie Everts-van der Wi, Hong-Seog Park, Jung-Sou Yeo, Inho Choi
Abstract
In this study, a Korean native cattle strain (Hanwoo) evidencing high performance in terms of both meat quality and quantity was employed in the generation of 150,000 BAC clones with an average insert size of 140 kb, and corresponding to about a 6X coverage of bovine chromosomal DNA. The BAC clones were pooled in a mini-scale via three rounds of a pooling protocol, and the efficiency of this pooling protocol was evaluated by testing the accuracy of accessibility to the positive clones, via a PCR-based screening method. Two sets of primers designed from each of two known genes were tested, and each yielded 2 or 3 positive clones for each gene, thereby indicating that the BAC library pooling system was appropriate with regard to the accession of the target BAC clones. Analyses of 3.3??06 base pairs obtained from the 7,090 BAC end sequence (BES) showed that 34.88% of the DNA sequence harbored the repetition sequence. Analysis of the 7,090 BES to the 1st and 2nd generation radiation hybrid map of the cattle genome, using the COMPASS program designed for the construction of a cattle-human comparative mapping, resulted in the localization of a total of 1,374 clones proximal to 339 1st generation markers, and 1,721 clones proximal to 664 2nd generation markers. Collectively, the BAC library and pooling system of the BAC clones from the Korean cattle, coupled with the chromosome-localized BAC clones, will provide us with novel tools for the excavation of desired clones for genome mapping and sequencing, and will also furnish us with additional information regarding breed differences in cattle.
Keywords: Bovine BAC Library; Pooling System; BAC End Sequence; Comparative Mapping; Chromosomal Localization


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