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.: Home > Animal Science Papers and Reports > 2004 > Volume 22 Number 4 > Alexey Sazanov1, Tadeusz Malewski2, Stanis³aw Kamiٌski3, Lech Zwierzchowski2

Identification of six BAC clones containing the bovine CSN1S1 and CSN3 genes: construction of the contig spanning the casein gene cluster*

Alexey Sazanov1, Tadeusz Malewski2, Stanis³aw Kamiٌski3, Lech Zwierzchowski2
1 Laboratory of Molecular Genome Organization, Institute of Farm Animal Genetics and Breeding, Russian Academy of Agricultural Sciences, Moskovskoye sh. 55A, St. Petersburg-Pushkin 196601, Russia 2 Department of Molecular Biology, Polish Academy of Sciences Institute of Genetics and Animal Breeding, Jastrzêbiec, 05-552 Wَlka Kosowska, Poland 3 Department of Animal Genetics, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, Micha³a Oczapowskiego 5, Poland
Abstract :

Regulatory sequences of the bovine casein genes are important for getting molecular markers of milk production traits and creation of high-effective vectors for transgenesis. Comparative analysis of these sequences requires continuous bovine DNA sequences covering the 5’-flanking region and coding part of the bovine casein gene cluster. Large insert clones containing CSN1S1 (لS1-casein) and CSN3 (ê-casein) genes with average insert size 180 kb were detected by screening of the bovine genomic BAC library CHORI-240 (6.1-fold genome coverage) with radioactively labelled DNAprobes specific for those genes. Six and thirteen BAC clones were identified by hybridization with CSN1S1- and CSN3-specific probes, respectively. The CH240-130F23 clone was confirmed to contain CSN1S1 gene by PCR amplification with specific primers; the CH240-046B08, CH240-115O19, CH240-258E02, and CH240-200N21 clones were shown to be specific for CSN3. The BAC clone CH240-035O12 was found to contain both casein genes as determined with PCR. These results show that the BAC contig (number of overlapping clones) was established for the whole casein cluster, including CSN1S1, CSN2, CSN1S2, and CSN3 genes. Based on the known insert sizes ofBAC clones in the CHORI-240 library, we conclude that this contig contains up to 270 kb of the 5’ flanking region and up to 270 kb of the 3’ flanking region. A maximum 5-fold coverage of the contig is assumed in the 3’ region of the cluster nearby the CSN3 gene.

Keywords :
BAC library / casein genes / cattle / clones / contig

Date Deposited : 11 Aug 2011 13:08

Last Modified : 11 Aug 2011 13:08

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Volume 22, Number 4, - 2004

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