Methylation of progesterone receptor isoform A and B promoters in the reproductive system of cows

2018 ◽  
Vol 30 (12) ◽  
pp. 1634 ◽  
Author(s):  
Robert Rekawiecki ◽  
Katarzyna Kisielewska ◽  
Magdalena K. Kowalik ◽  
Jan Kotwica

The aim of this study was to investigate whether the promoters of progesterone receptor isoform A (PGRA) and B (PGRB) are methylated and to determine the percentage of methylation occurring for each isoform. Genomic DNA was isolated from the corpora lutea (CL) and endometrial slices from cows on Days 2–5, 6–10, 11–16 and 17–20 of the oestrous cycle. DNA was bisulphite-converted and amplified using methyl-specific polymerase chain reaction (PCR) with primers that detect both methylated and unmethylated sequences. The determination of the percentage of the methylation was performed using HpaII and MspI restriction enzymes. Methyl-specific PCR showed partial methylation of PGRA and PGRB promoters in the CL and endometrium during the oestrous cycle. Methylation for PGRA was between 15 and 17% and for PGRB was in the range of 6 to 7.7% during the oestrous cycle in the CL. In the endometrium, the methylation for PGRA was between 6 and 7.3% and for PGRB was between 3 and 4.8% during the oestrous cycle. The data obtained indicate that the higher promoter methylation of the PGRA isoform could be a mechanism for regulation of PGRA inhibitory activity against PGRB and, in this way, methylation may influence the regulation of progesterone action in the CL and endometrium.

Transfusion ◽  
1995 ◽  
Vol 35 (3) ◽  
pp. 231-240 ◽  
Author(s):  
D. F. Stroncek ◽  
R. Konz ◽  
M. E. Clay ◽  
J. P. Houchins ◽  
J. McCullough

2014 ◽  
Vol 14 (2) ◽  
pp. 10-15 ◽  
Author(s):  
Mendelova A. ◽  
Holubekova V. ◽  
Zubor P. ◽  
Jezkova E. ◽  
Gemzova K. ◽  
...  

Abstract The phosphatidylinositol 3-kinases (PI3Ks) are a family of proteins involved in the regulation of cell survival, growth, metabolism, and glucose homeostasis. Increased PI3K activity is associated with many cancers. PIK3CA gene (encoding p110 , the catalytic subunit of PI3K) is commonly mutated in breast cancer. In our study we focused on the detection of “hotspot” mutations in exons 9 and 20 of the PIK3CA gene in paraffin-embedded tissue of patients with breast cancer. We optimized conditions of allele specific polymerase chain reaction (PCR) and we used direct sequencing to verify our results. Overall, three “hotspot” mutations in PIK3CA gene in paraffin-embadded tissue from breast cancer were detected by allele-specific PCR. All results were verified by direct sequencing of PCR products and we observed 100% agreement between those two methods. We confirmed that allele-specific PCR assay is low cost method usefull for accurate detection of PIK3CA mutations.


Author(s):  
Ahmed M. Darwish ◽  
Ghada H. H. El Nady ◽  
Neama I. Ali ◽  
Aly Z. E. Abdelsalam

The beta casein gene (CSNS2) has 12 genetic variants divided into two groups: the first group (A1, B, and G) which differ from the second group (A2, A3 C, D, E, F, H, H2 and I) where A base replaces C base, this leads to potential liberation of a bioactive peptide, b-casomorphin, upon digestion where a histidine replaces a proline at position 67. The allele specific polymerase chain reaction (AS-PCR) was evaluated to distinguish between the beta casomorphin releasing variants (A1 and B) and the non-releasing variants. The sequence analysis was used to determine these variants and confirm it in goat, sheep and cattle. The results showed that cattle carrying allele A1 either homozygous or heterozygous more than sheep and goats. The allele frequency of A1 and A2 is 0.44, 0.56 in goats, 0.43, 0.57 in sheep and 0.54, 0.46 in cattle, respectively. The sequence results reported changing of C base to A base in goat, sheep and cattle. Therefore, this study reported that goat and sheep milk was more safe than cattle milk.


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