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Titolo:
AN INDELIBLE LINEAGE MARKER FOR XENOPUS USING A MUTATED GREEN FLUORESCENT PROTEIN
Autore:
ZERNICKAGOETZ M; PINES J; RYAN K; SIEMERING KR; HASELOFF J; EVANS MJ; GURDON JB;
Indirizzi:
WELLCOME CRC INST,TENNIS COURT RD CAMBRIDGE CB2 1QR ENGLAND UNIV CAMBRIDGE,DEPT ZOOL,MRC,MOL BIOL LAB CAMBRIDGE CB2 1TN ENGLAND UNIV CAMBRIDGE,DEPT GENET,MRC,MOL BIOL LAB CAMBRIDGE CB2 1TN ENGLAND
Titolo Testata:
Development
fascicolo: 12, volume: 122, anno: 1996,
pagine: 3719 - 3724
SICI:
0950-1991(1996)122:12<3719:AILMFX>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
CENTRAL NERVOUS-SYSTEM; CELL LINEAGE; INDIVIDUAL BLASTOMERES; CLONAL ORGANIZATION; 32-CELL STAGE; CHICK-EMBRYO; GENE; EXPRESSION; DROSOPHILA; LAEVIS;
Keywords:
XENOPUS; GREEN FLUORESCENT PROTEIN; MARKER; EOMES; CELL LINEAGE; GFP.RN3;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
39
Recensione:
Indirizzi per estratti:
Citazione:
M. Zernickagoetz et al., "AN INDELIBLE LINEAGE MARKER FOR XENOPUS USING A MUTATED GREEN FLUORESCENT PROTEIN", Development, 122(12), 1996, pp. 3719-3724

Abstract

We describe the use of a DNA construct (named GFP.RN3) encoding greenfluorescent protein as a lineage marker for Xenopus embryos. This offers the following advantages over other lineage markers so far used inXenopus. When injected as synthetic mRNA, its protein emits intense fluorescence in living embryos. It is non-toxic, and the fluorescence does not bleach when viewed under 380 nm light, It is surprisingly stable, being strongly visible up to the feeding tadpole stage (5 days), and in some tissues for several weeks after mRNA injection. We also describe a construct that encodes a blue fluorescent protein. We exemplify the use of this GFP.RN3 construct for marking the lineage of individual blastomeres at the 32- to 64-cell stage, and as a marker for single transplanted blastula cells. Both procedures have revealed that the descendants of one embryonic cell can contribute single muscle cells to nearly all segmental myotomes rather than predominantly to any one myotome. An independent aim of our work has been to follow the fate of cells in which an early regulatory gene has been temporarily overexpressed, For this purpose, we co-injected GFP.RN3 mRNA and mRNA for the early Xenopus gene Eomes, and found that a high concentration of Eomes results in ectopic muscle gene activation in only the injected cells. This marker may therefore be of general value in providing long term identification of those cells in which an early gene with ephemeral expression has been overexpressed.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 01/12/20 alle ore 08:00:56