The complete set of predicted genes from Saccharomyces cerevisiae in a readily usable form

Jr Hudson, E. P. Dawson, K. L. Rushing, C. H. Jackson, D. Lockshon, D. Conover, C. Lanciault, J. R. Harris, S. J. Simmons, R. Rothstein, S. Fields

Research output: Contribution to journalArticlepeer-review

111 Scopus citations

Abstract

Nearly all of the open reading frames (ORFs) of the yeast Saccharomyces cerevisiae have been synthesized by PCR using a set of ~6000 primer pairs. Each of the forward primers has a common 22-base sequence at its 5' end, and each of the back primers has a common 20-base sequence at its 5' end. These common termini allow reamplification of the entire set of original PCR products using a single pair of longer primers - in our case, 70 bases. The resulting 70-base elements that flank each ORF can be used for rapid and efficient cloning into a linearized yeast vector that contains these same elements at its termini. This cloning by genetic recombination obviates the need for ligations or bacterial manipulations and should permit convenient global approaches to gene function that require the assay of each putative yeast gene.

Original languageEnglish (US)
Pages (from-to)1169-1173
Number of pages5
JournalGenome Research
Volume7
Issue number12
DOIs
StatePublished - 1997
Externally publishedYes

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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