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

Yeast. 2000 Dec;16(16):1527-34. doi: 10.1002/1097-0061(200012)16:16<1527::AID-YEA643>3.0.CO;2-8.

Abstract

Creation of large ( approximately 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 I intron ribozyme from Candida albicans. This plasmid expresses functional rRNA and ribozyme.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Candida albicans / genetics
  • Genetic Engineering / methods*
  • Plasmids*
  • Polymerase Chain Reaction
  • RNA, Catalytic / genetics
  • RNA, Fungal / genetics
  • RNA, Ribosomal / genetics
  • Recombination, Genetic
  • Saccharomyces cerevisiae / genetics*
  • Transformation, Genetic

Substances

  • RNA, Catalytic
  • RNA, Fungal
  • RNA, Ribosomal
  • RNA, ribosomal, 25S