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Our earlier studies carried out in silico demonstrated that different transcription factors have their putative binding sites in the 5’-flanking regions of rabbit milk protein genes. Now we extended his study to include the experimental analysis of the transcription factors. This study of electrophoretic mobility shift assay (EMSA) of nuclear proteins showed for the first time the presence of AP1, CREB, Myb, and Sp1 transcription factors in the rabbit mammary gland. The abundance of all transcription factors studied in the mammary gland changed during pregnancy-lactation-weaning cycle. The DNA-binding activity of the Myb transcription factor correlated well with the level of milk protein gene expression. The affinity of Myb to its binding sequence is methylation-dependent. One of CREspecific DNA-protein complexes was found only in nuclear protein from the lactating rabbits mammary gland. The amount of AP1 DNA-binding activity transiently decreased in the early involution. Transition from early to late involution was associated with the restoration of the AP1 activity, and with the decrease of the Myb and Sp1 protein-DNA binding activities.
Our earlier studies carried out in silico demonstrated that different transcription factors have their putative binding sites in the 5’-flanking regions of bovine milk protein genes. Now we extended our study to include the experimental analysis of these transcription factors. This study on electrophoretic mobility shift assay (EMSA) of nuclear proteins derived from bovine mammary glands showed for the first time the presence there of CREB, NF-κB, Oct1 and Sp1 transcription factors. The pattern of DNA-protein complexes of Sp1 transcription factor significantly differed between virgin heifers and lactating cows. Transition from pregnancy to lactation was associated with changes in the DNAprotein binding patterns of NFI and Oct1 transcription factors.
The effects of cow’s genotype at αS1- and αS2-casein gene 5’-noncoding regions (promoters) were determined on selected milk production traits of the 135 Polish Black-and-White (Polish Friesian) cows as related to the animal’s age, lactation parity and stage, and somatic cell count. Cows of the AA genotype of αS1-casein gene yielded more milk daily than AG heterozygotes. Also, the daily yield of solids-non-fat, protein and lactose was higher in AA genotype cows. Milk of the cows with genotype CC of αS2-casein contained more lactose but less protein than that of the CT heterozygotes. The daily protein yield was slightly (but significantly) higher in the cows of the CT αS2-casein genotype. In summary, the results showed that genetic variants of αS1- and αS2-casein 5’-noncoding regions had only a slight effect on milk production traits of the Polish Black-and-White cows. Nevertheless,the AA genotype of αS1-casein seemed favourable for higher milk yield, as well as for lactose and protein content.
Nineteen ВАС clones were identified by hybridization of the bovine genomic ВАС library CHORI-240 with mixed CSN1S1- and CSN3-specific probes. Two of the clones were shown to contain the genes CSN1S1, CSN1S2, CSN2, STATH and CSN3, and five were proved to include the genes CSN2, STATH, CSN1S2 and CSN3. These data showed that the ВАС contig was established for the whole casein cluster, including all known five genes.
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