DETECTION AND MOLECULAR CLONING OF GROWTH DIFFERENTIATION FACTOR-9 TRANSCRIPTS IN IN VITRO MATURED BUFFALO OOCYTE COMPLEXES

Article Id: ARCC180 | Page : 472-478
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
H.N. Malik*, S.K. Dash1, S.K. Mohapatra2 and T. Nanda hrudananda.malik@gmail.com
Address : Animal Biotechnology Center, CCS Haryana Agricultrual University, Hisar-125 004, India

Abstract

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.

Keywords

Cumulus oocyte complexes Follicle In vitro maturation.

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