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 journalArticlepeer-review

1 Scopus citations

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

ASJC Scopus subject areas

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

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