MOLECULAR CHARACTERIZATION OF CYTOCHROME P450 AROMATASE (CYP19) GENE IN EGYPTIAN RIVER BUFFALOES
Abstract
Cytochrome P450 aromatase (CYP19) gene catalyzes the last step in the steroidogenesis which convert the androgens to estrogens. Therefore, the objective of this study was to investigate the polymorphism in CYP19 gene and its potential effects in female buffaloes fertility. The DNA was extracted from the blood samples of 81 Egyptian river buffalo females and a 419 bp fragment contained a part of CYP19- distal promoter P1.1 was amplified by PCR which subsequently treated with PvuII restriction enzyme. The PCR-RFLP pattern showed that all the animals had a fixed GG genotype and A allele was not detected. Sequencing of the amplified fragment (GenBank accession No. MF490278 and MF490279) followed by sequence alignment with the GenBank database revealed that the homology among the Egyptian river buffalo target sequence and its orthologues sequences in cattle, sheep and goat was 97, 95 and 93%, respectively. G to A transition SNP (G197A) was detected among individuals representing the Egyptian river buffalo by sequencing. G allele was detected only in the Egyptian buffalo and not in the other buffalo records in the GenBank. Seventeen transcription factor binding sites (TFBSs) span along the sequence were predicted.References
Blakemore, J. and F. Naftolin (2016). Aromatase: contributions to physiology and disease in women and men. Physiology (Bethesda), 31: 258-269.
Di Nardo, G. and G. Gilardi (2013). Human aromatase: perspectives in biochemistry and biotechnology. Biotechnol. Appl. Biochem., 60: 92-101.
Grabe, N. (2002). AliBaba2: context specific identification of transcription factor binding sites. In Silico Biol., 2: S1-S15.
Iannuzzi, L., G. P. Di Meo, A. Perucatti, L. Schibler, D. Incarnato and E. P. Cribiu (2001). Comparative FISH mapping in river buffalo and sheep chromosomes: assignment of forty autosomal type I loci from sixteen human chromosomes. Cytogenet. Cell Genet., 94: 43-48.
Miller, S. A., D. D. Dykes and H. F. Polesky (1988). A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res., 16: 1215.
Perera, B. M. A. O. (2011). Reproductive cycles of buffalo. Anim. Reprod. Sci., 124: 194-199.
Szatkowska, I., M. Jedrzejczak, A. Dybus, W. Grzesiak, D. Zaborski and D. Jankowiak (2011). An analysis of CYP19, CYP21 and ER genotypes in Polish Holstein-Friesian cows with regard to the selected reproductive traits. Acta. Vet. Brno., 80: 65-71.
Tantia, M. S., R. K. Vijh, P. K. Vij, R. S. Kataria, B. P. Mishra, B. K. Joshi, V. Bhasin, S. C. Gupta, K. M. L. Pathak, P. Sikka, S. P. Yadav, A. K. Pandey and R. K. Sethi (2011). Whole-genome sequence assembly of the water buffalo (Bubalus bubalis). Indian Journal of Animal Sciences, 81: 38-46.
Wierzbicki, H., E. Bula and M. Kmiec (2014). Polymorphism of cytochrome P450 aromatase gene and its association with selected reproductive traits in Polish Holstein-Friesian cattle. J. Anim. Vet. Adv., 13: 1098-1103.
Yazdani, M. S. and S. Ahmed (2010). An "on the spot" test for targeted screening in index families of thalassaemia. J. Pak. Med. Assoc., 60: 521-523.
Yazdani, S. K., M. Nakano, F. Otsuka, F. D. Kolodgie and R. Virmani (2010). Atheroma and coronary bifurcations: before and after stenting. Euro Intervention, 6 Suppl J: J24-30.
