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Titolo:
Comparison by restriction fragment differential display RT-PCR of gene expression pattern in bovine oocytes matured in the presence or absence of fetal calf serum
Autore:
Rzucidlo, SJ; Gibbons, J; Stice, SL;
Indirizzi:
Univ Georgia, Dept Anim & Dairy Sci, Athens, GA 30602 USA Univ Georgia Athens GA USA 30602 t Anim & Dairy Sci, Athens, GA 30602 USA Prolinia Inc, Athens, GA USA Prolinia Inc Athens GA USAProlinia Inc, Athens, GA USA
Titolo Testata:
MOLECULAR REPRODUCTION AND DEVELOPMENT
fascicolo: 1, volume: 59, anno: 2001,
pagine: 90 - 96
SICI:
1040-452X(200105)59:1<90:CBRFDD>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
IN-VITRO MATURATION; DEVELOPMENTAL COMPETENCE; LUTEINIZING-HORMONE; PROTEIN-SYNTHESIS; MESSENGER-RNA; PHOSPHORYLATION PATTERNS; INVITRO FERTILIZATION; EMBRYONIC-DEVELOPMENT; CULTURE; MEDIA;
Keywords:
mRNA; cDNA; differential gene expression; in vitro maturation; bovine oocytes;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
38
Recensione:
Indirizzi per estratti:
Indirizzo: Stice, SL Univ Georgia, Dept Anim & Dairy Sci, Athens, GA 30602 USA Univ Georgia Athens GA USA 30602 airy Sci, Athens, GA 30602 USA
Citazione:
S.J. Rzucidlo et al., "Comparison by restriction fragment differential display RT-PCR of gene expression pattern in bovine oocytes matured in the presence or absence of fetal calf serum", MOL REPROD, 59(1), 2001, pp. 90-96

Abstract

A novel restriction fragment differential display (RFDD) RT-PCR has been used to compare patterns of mRNA expression in bovine oocytes matured in vitro in the presence (10%) or absence of fetal calf serum (FCS). Total RNA extracted from matured and denuded oocytes was processed using displayProfilekit (Display System Biotech). RFDD RT-PCR products were separated on 6% polyacrylamide gel and analyzed using a Storm 860 scanner. Selected bands representing potentially differentially expressed fragments were excised from the gel and re-amplified. Re-amplified fragments with size matched to the original fragment were cloned into the TA vector and sequenced. Initially, 10 and 15 differentially expressed fragments were isolated from oocytes matured in the presence and absence of FCS, respectively. Eight out of 10 and 10 out of 15 fragments were re-amplified successfully as evidenced by size similarity to the original fragments. Finally, the size of six inserts sequenced from each group matched the size of corresponding original as well as re-amplified fragments. Sequence comparison search revealed similarity of some isolated fragments to 18s ribosomal RNA, bovine apolipoprotein A-I, bovine mitochondrion DNA, human CGI-79 mRNA, human Abl-interactor protein, andbovine satellite DNA. The other sequenced fragments may represent novel genes. We showed that RFDD RT-PCR can be effectively applied to contrast geneexpression pattern in bovine oocytes and that presence or absence of FCS during maturation interval affects gene expression pattern in matured bovineoocytes. Mol. Reprod. Dev. 59:90-96, 2001. (C) 2001 Wiley-Liss, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 07/08/20 alle ore 00:30:00