Disruption and phenotypic analysis of six open reading frames from chromosome V...

• Autor(es): Guerreiro, Paulo ; Rodrigues-Pousada, Claudina
• Data: 2001
• Identificador Persistente: http://hdl.handle.net/10400.21/3086
• Origem: Repositório Científico do Instituto Politécnico de Lisboa
• Assunto(s): Saccharomyces cerevisiae; EUROFAN; Gene disruption; ZPR1; YAP1802; LSC2; YHR162w; Chromosomes; Gene deletion; Mutagenesis, Insertional; Open reading frames/genetics; Phenotype; Plasmids/genetics; Polymerase chain reaction; Transformation, Genetic
• Tipo de Documento: Artigo
• Idioma: Inglês

Descrição

Six open reading frames (ORFs) located on chromosome VII of Saccharomyces cerevisiae (YGR205w, YGR210c, YGR211w, YGR241c, YGR243w and YGR244c) were disrupted in two different genetic backgrounds using short-flanking homology (SFH) gene replacement. Sporulation and tetrad analysis showed that YGR211w, recently identified as the yeast ZPR1 gene, is an essential gene. The other five genes are non-essential, and no phenotypes could be associated to their inactivation. Two of these genes have recently been further characterized: YGR241c (YAP1802) encodes a yeast adaptor protein and YGR244c (LSC2) encodes the b-subunit of the succinyl-CoA ligase. For each ORF, a replacement cassette with long flanking regions homologous to the target locus was cloned in pUG7, and the cognate wild-type gene was cloned in pRS416.