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Titolo:
THE USE OF A DOUBLE-MARKER SHUTTLE VECTOR TO STUDY DNA DOUBLE-STRAND BREAK REPAIR IN WILD-TYPE AND RADIATION-SENSITIVE MUTANTS OF THE YEASTSACCHAROMYCES-CEREVISIAE
Autore:
JHA B; AHNE F; ECKARDTSCHUPP F;
Indirizzi:
L N MITHILA UNIV,DEPT BOT DARBHANGA 846004 INDIA L N MITHILA UNIV,DEPT BOT DARBHANGA 846004 INDIA GSF MUNICH,FORSCHUNGSZENTRUM UMWELT & GESUNDHEIT,INST STRAHLENBIOL W-8042 NEUHERBERG GERMANY
Titolo Testata:
Current genetics
fascicolo: 5-6, volume: 23, anno: 1993,
pagine: 402 - 407
SICI:
0172-8083(1993)23:5-6<402:TUOADS>2.0.ZU;2-T
Fonte:
ISI
Lingua:
ENG
Soggetto:
FIELD GEL-ELECTROPHORESIS; IONIZING-RADIATION; GENETIC-CONTROL; ATAXIA-TELANGIECTASIA; DEOXYRIBONUCLEIC-ACID; ESCHERICHIA-COLI; MAMMALIAN-CELLS; GAP REPAIR; URA3 GENE; RECOMBINATION;
Keywords:
DOUBLE-MARKER PLASMID; DNA DOUBLE-STRAND BREAK; YEAST TRANSFORMATION; FIDELITY OF REPAIR;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
48
Recensione:
Indirizzi per estratti:
Citazione:
B. Jha et al., "THE USE OF A DOUBLE-MARKER SHUTTLE VECTOR TO STUDY DNA DOUBLE-STRAND BREAK REPAIR IN WILD-TYPE AND RADIATION-SENSITIVE MUTANTS OF THE YEASTSACCHAROMYCES-CEREVISIAE", Current genetics, 23(5-6), 1993, pp. 402-407

Abstract

An episomal DNA vector (YpJA18), encoding two selectable recombinant yeast genes (TRP1, URA3), was constructed to assess the fidelity of DNA repair in haploid repair-competent (RAD) wild-type yeast and severalradiation-sensitive mutants. Either a DNA double-strand break (DSB) or a double-strand gap of 169 bp (DSG) was introduced by restriction enzymes in-vitro within the coding sequence of the URA3 gene of this vector. To eliminate transfer artefacts, selection was first applied for the undamaged TRP1 gene followed by counter selection for URA3 gene activity, which indicated correct repair of the DSB and DSG. Correct repair of the damaged URA3 gene was found to be about 90 % in RAD cells (normalized for the expression of undamaged URA3 in TRP+ transformants). Plasmids isolated from the transformants (URA+TRP+) carry both unique sites (ApaI and NcoI) within the URA3 gene indicating the precise restitution of the 169-bp gap. An excision-repair-defective rad4-4 mutant repaired these lesions as correctly as RAD cells, whereas the mutants rad50-1, rad51-1 and rad54-1, proven to be defective in DSB repair and mitotic recombination, showed less than 5 % correct repair of such lesions. In contrast, a representative of the RAD6 epistasis group of genes, the rev2-1 mutant which is sensitive towards UV and ionizing radiation, had a significantly reduced ability (about 20 %) for the correct repair of both DSBs and DSGs.

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Documento generato il 24/09/20 alle ore 04:52:49