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Identification of some unknown transcripts from SSH cDNA library of buffalo follicular oocytes

Published online by Cambridge University Press:  10 August 2012

S. K. Rajput ,
P. Kumar ,
B. Roy ,
A. Verma ,
H. P. Pandey ,
D. Singh ,
S. De  and
T. K. Datta
S. K. Rajput
Affiliation:
National Dairy Research Institute, Animal Biotechnology Centre, Karnal 132001, India
P. Kumar
Affiliation:
National Dairy Research Institute, Animal Biotechnology Centre, Karnal 132001, India
B. Roy
Affiliation:
National Dairy Research Institute, Animal Biotechnology Centre, Karnal 132001, India
A. Verma
Affiliation:
National Dairy Research Institute, Animal Biotechnology Centre, Karnal 132001, India
H. P. Pandey
Affiliation:
Department of Biochemistry, Banaras Hindu University, 221005 Varanasi, India
D. Singh
Affiliation:
National Dairy Research Institute, Animal Biotechnology Centre, Karnal 132001, India
S. De
Affiliation:
National Dairy Research Institute, Animal Biotechnology Centre, Karnal 132001, India
T. K. Datta*
Affiliation:
National Dairy Research Institute, Animal Biotechnology Centre, Karnal 132001, India
*
†E-mail: tirthadatta@gmail.com
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Abstract

A buffalo oocyte-specific subtracted cDNA library was constructed to identify exclusively or preferentially oocyte-expressed genes. The library represented an enriched population of transcripts obtained from oocytes of diverse ovarian follicular origin and at different stages of in vitro maturation. A total of 1173 high-quality sequences of oocyte-specific genes were clustered into 645 unique sequences, out of which 65.76% were represented as singlets and 34.26% as contig expressed sequence tags (ESTs; clusters). Analysis of sequences revealed that 498 of these sequences were identified as a known sequence in mammalian species including buffalo, 103 as uncharacterized ESTs and 44 unknown sequences including 1 novel EST, so far not reported in any species. Gene ontology annotation classified these sequences into functional categories of cellular events and biological processes associated with oocyte competence. Expression status of the isolated unknown ESTs confirmed that many of these are expressed in oocytes exclusively and in others preferentially, some in excess of 80-fold greater in comparison with a variety of somatic tissues. The isolated novel EST was detected to be expressed exclusively in oocytes and testicular cells only. To our knowledge, this is the first report giving a detailed transcriptome account of oocyte-expressed genes in buffalo. This study will provide important information on the physiological control of oocyte development, as well as many questions yet to be addressed on the reproductive process of buffalo.

Keywords

buffalooocytecDNA libraryEST
Type
Physiology and functional biology of systems
Information
animal , Volume 7 , Issue 3 , March 2013 , pp. 446 - 454
Copyright
Copyright © The Animal Consortium 2012

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