• Title/Summary/Keyword: BAC End Sequence (BES)

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Localization of 5,105 Hanwoo (Korean Cattle) BAC Clones on Bovine Chromosomes by the Analysis of BAC End Sequences (BESs) Involving 21,024 Clones

  • Choi, Jae Min;Chae, Sung-Hwa;Kang, Se Won;Choi, Dong-Sik;Lee, Yong Seok;Park, Hong-Seog;Yeo, Jung-Sou;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.11
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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.

Chromosomal Localization of Korean Cattle (Hanwoo) BAC Clones via BAC end Sequence Analysis

  • Chae, Sung-Hwa;Kim, Jae-Woo;Choi, Jae Min;Larkin, Denis M.;Everts-van der Wind, Annelie;Park, Hong-Seog;Yeo, Jung-Sou;Choi, Inho
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.3
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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.

The 3rd Generation Genome Map of the Korean Cattle (Hanwoo) (제3세대 한우유전체지도작성)

  • Lee, Yong-Seok;Choi, In-Ho
    • Journal of Animal Science and Technology
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    • v.51 no.2
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    • pp.123-128
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    • 2009
  • Recently, the $2^{nd}$ generation genome map of the Korean cattle (Hanwoo) has been constructed by comparison of the nucleotide sequence of the Korean cattle BAC clones with whole genome sequence of the bovine data-base (B_tau 2.1 build). The objective of this study was to update the $2^{nd}$ generation genome map of the Korean cattle using the similar approach. The nucleotide sequence of the Korean cattle BAC clones utilized in the construction of the $2^{nd}$ generation map was compared with the newly released bovine data-base (B_tau 3.1 build) to generate the $3^{rd}$ generation map. While, 5,105 BAC clones were localized on bovine chromosome in the $2^{nd}$ generation map, a total of 9,595 BAC clones, which spans about 37.27% of the bovine chromosome after eliminating the overlapping sequence among the clones, have been mapped on the bovine chromosome in the $3^{rd}$ generation map. Further analysis of the nucleotide sequence of the BAC clones will allow us to develop map and facilitate to pinpoint the genes that are important for the improvement of the performance in this cattle breed.