• Asian-Australasian Journal of Animal Sciences
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• 제20권11호
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• pp.1636-1650
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• 2007

As an initial step toward a better understanding of the genome structure of Korean cattle (Hanwoo breed) and initiation of the framework for genomic research in this bovine, the bacterial artificial chromosome (BAC) end sequencing of 21,024 clones was recently completed. Among these clones, BAC End Sequences (BESs) of 20,158 clones with high quality sequences (Phred score ${\geq}20$, average BES equaled 620 bp and totaled 23,585,814 bp), after editing sequencing results by eliminating vector sequences, were used initially to compare sequence homology with the known bovine chromosomal DNA sequence by using BLASTN analysis. Blast analysis of the BESs against the NCBI Genome database for Bos taurus (Build 2.1) indicated that the BESs from 13,201 clones matched bovine contig sequences with significant blast hits (E<$e^{-40}$), including 7,075 single-end hits and 6,126 paired-end hits. Finally, a total of 5,105 clones of the Korean cattle BAC clones with paired-end hits, including 4,053 clones from the primary analysis and 1,052 clones from the secondary analysis, were mapped to the bovine chromosome with very high accuracy.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.316-327
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• 2007

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 $3.3{\times}10^6$ 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 $1^{st}$ and $2^{nd}$ 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 $1^{st}$ generation markers, and 1,721 clones proximal to 664 $2^{nd}$ 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.

• Journal of Animal Science and Technology
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• 제51권2호
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• pp.123-128
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• 2009

최근 한우의 유전체 연구의 핵심 소재인 박테리아인공염색체(BAC; Bacterial Artificial Chromosome) 클론이 제작되었고 이에 대한 염기서열의 해독과 이를 NCBI (National Center for Biotechnology Information)의 소 유전체 데이터(B_tau 2.1)와 비교 분석하여 한우의 유전체지도 초안(제2 세대 한우유전체지도)이 제작되었다. 본 연구에서는 기존에 확보한 한우의 박테리아인공염색체 클론의 염기서열을 최근에 공개된 소유전체 데이터(B-tau 3.1)와 비교 분석하여 한우 유전체 지도를 최신화하기 위해 실시하였다. 제2 세대 한우유전체지도에서 총 5,105개의 클론에 대한 염색체상 위치가 결정된 반면에 제3세대 한우유전체지도에서는 총 9,595개의 클론에 대한 염색체 위치가 결정되어 약 2배 정도로 향상되었다. 또한 겹치는 부분을 제외했을 때 제3세대 한우유전체지도에 사용된 클론은 소 전체 염색체의 약 37.27%에 해당되는 부분을 커버하는 것으로 나타났다. 앞으로 추가적인 한우 클론의 염기서열 해독을 통해 얻어진 데이터를 확보한다면 한우염색체 정밀 지도의 완성과 이를 이용한 한우 개량에 유용한 유전자 발굴에 활용될 수 있을 것으로 판단되다.