Construction of a large plasmid lacking linearizing single restriction sites by simultaneous in vivo recombination and plasmid shuffling in yeast

Karl E. Miletti, Michael J Leibowitz

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Creation of large (∼ 15 kb) recombinant plasmids can be done in a single step by co-transformation of yeast cells with a partial restriction digest of a plasmid vector and a linear insert whose ends overlap one of the vector restriction sites. This method is used to generate a plasmid expressing the Saccharomyces cerevisiae rRNA genes containing the Ca.LSU group 1 intron ribozyme from Candida albicans. This plasmid expresses functional rRNA and ribozyme.

Original languageEnglish (US)
Pages (from-to)1527-1534
Number of pages8
JournalYeast
Volume16
Issue number16
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Yeast
Genetic Recombination
plasmids
Plasmids
Yeasts
yeasts
Catalytic RNA
Candida
ribosomal RNA
plasmid vectors
Genes
Cells
Candida albicans
introns
Saccharomyces cerevisiae
rRNA Genes
Introns
genes
cells
methodology

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Microbiology

Cite this

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