Yeast vectors for the integration/expression of any sequence at the TYRI locus

Mario Giuseppe Mirisola, Giacomo De Leo, Mario G. Mirisola, Alessia Gallo, Rosa Maria Amodeo

Risultato della ricerca: Articlepeer review

2 Citazioni (Scopus)

Abstract

We have constructed new yeast vectors for targeted integration and conditional expression of any sequence at the Saccharomyces cerevisiae TYR1 locus which becomes disrupted. We show that vector integration is not neutral, causing prototrophy for tyrosine and auxotrophy for the vector's selectable marker (uracil or leucine, depending on the vector used). This feature allows a double screening of transformed yeast cells, improving the identification of colonies with the desired chromosomal structure. The GAL10 gene promoter has been added to drive conditional expression of cloned sequences. Using these vectors, chromosomal structure verification of recombinant clones is no longer necessary, since the noise of non-homologous recombination, as well as spontaneous reversion of the selected phenotype, can easily be identified. The ability of the vector to conditionally control gene expression has been confirmed using the gene for the green fluorescent protein (GFP) as a reporter.
Lingua originaleEnglish
pagine (da-a)761-766
Numero di pagine6
RivistaYeast
Volume24
Stato di pubblicazionePublished - 2007

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Genetics

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