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Titolo:
Single strand mRNA differential display (SSDD) applied to the identification of serine/threonine phosphatases regulated during cerebellar development
Autore:
Vila-Ortiz, GJ; Radrizzzani, M; Carminatti, H; Idoyaga-Vargas, VP; Santa-Coloma, TA;
Indirizzi:
CONICET, IIBBA, UBA,IIB, Fdn Campomar,Inst Invest Bioquim, RA-1405 Buenos Aires, DF, Argentina CONICET Buenos Aires DF Argentina RA-1405 405 Buenos Aires, DF, Argentina Inst Malbran, ANLIS, Ctr Nacl Genet Med, Malbran, Argentina Inst Malbran Malbran Argentina , Ctr Nacl Genet Med, Malbran, Argentina
Titolo Testata:
JOURNAL OF NEUROSCIENCE METHODS
fascicolo: 1, volume: 105, anno: 2001,
pagine: 87 - 94
SICI:
0165-0270(20010130)105:1<87:SSMDD(>2.0.ZU;2-R
Fonte:
ISI
Lingua:
ENG
Soggetto:
SERINE THREONINE PHOSPHATASES; POLYMERASE CHAIN-REACTION; PROTEIN PHOSPHATASES; MESSENGER-RNA; POLYACRYLAMIDE GELS; PAPILLARY CARCINOMA; DNA; EXPRESSION; BRAIN; CDNA;
Keywords:
single strand differential display; differential display; single strand conformation polymorphism; serine threonine phosphatases; development; cerebellum;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Santa-Coloma, TA CONICET, IIBBA, UBA,IIB, Fdn Campomar,Inst Invest Bioquim, Patricias Argentinas 435, RA-1405 Buenos Aires, DF, Argentina CONICET Patricias Argentinas 435 Buenos Aires DF Argentina RA-1405
Citazione:
G.J. Vila-Ortiz et al., "Single strand mRNA differential display (SSDD) applied to the identification of serine/threonine phosphatases regulated during cerebellar development", J NEUROSC M, 105(1), 2001, pp. 87-94

Abstract

Differential display is a used widely and useful technique for the study of differentially expressed genes. However, very poor results have been obtained in the past when particular gene families were studied. Initially, we attempted to study the mRNA expression of catalytic subunits of serine/threonine phosphatases, using two primers specific to consensus sequences of these phosphatases. When differential display was applied, two wide, unresolved bands were isolated that contained cDNA of several phosphatases. together with that of many other unrelated transcripts. To overcome this problem, we used an alternative strategy, referred to as single strand differential display (SSDD), which is a combination of differential display and single strand conformation polymorphism (SSCP). After initial PCR amplification with specific primers, we ran a polyacrylamide (or agarose) gel, pre-selectingthe region that contained fragments of the size expected for the consensusregion (250-350 bp). The DNA eluted from this zone was then separated on anon-denaturing (SSCP) gel. Using this approach, we were able to characterize the expression of five ser/thr phosphatases, and a previously unreportedsplice variant of one of them, PP1 gamma. All these phosphatases show varying levels of expression during development, indicating a very complex regulation of protein phosphorylation-dephosphorylation during the period of synaptogenesis in the mouse cerebellum. (C) 2001 Elsevier Science B.V. All rights reserved.

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Documento generato il 04/04/20 alle ore 08:45:57