DETECTION AND MOLECULAR CLONING OF GROWTH DIFFERENTIATION FACTOR-9 TRANSCRIPTS IN IN VITRO MATURED BUFFALO OOCYTE COMPLEXES
Citation :- DETECTION AND MOLECULAR CLONING OF GROWTH DIFFERENTIATION FACTOR-9 TRANSCRIPTS IN IN VITRO MATURED BUFFALO OOCYTE COMPLEXES.Indian Journal Of Animal Research.2013.(47):472-478
The present study was carried out on buffalo ovarian follicles. From these follicles cumulus oocyte complexes were aspirated and in vitro matured in TCM-199 supplemented 10% v/v fetal bovine serum, 1µg/ml estradiol, 10µg/ml LH, 5µg/ml FSH and 1% v/v gentamycin. In vitro matured cumulus oocyte complexes were investigated for the presence of GDF -9 gene transcript using RT-PCR analysis. After the RT-PCR analysis the amplified product was purified and cloned. The cloned product was further sequenced and analyzed to study the phylogenetic relationship of GDF-9 gene of buffalo with other species. Upon RT-PCR analysis it was revealed that GDF-9 transcripts were expressed exclusively in in vitro matured cumulus oocyte complexes. Nucleotide sequence analysis of GDF-9 gene of Bubalus bubalis revealed that it is 99% similar to that of Bos taurus and clustered together with Bos taurus and Rattus norvegicus shared a separate cluster.
Cumulus oocyte complexes
In vitro maturation.
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