Premium
Construction of a large plasmid lacking linearizing single restriction sites by simultaneous in vivo recombination and plasmid shuffling in yeast
Author(s) -
Miletti Karl E.,
Leibowitz Michael J.
Publication year - 2000
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/1097-0061(200012)16:16<1527::aid-yea643>3.0.co;2-8
Subject(s) - biology , shuffling , plasmid , genetics , recombination , yeast , dna shuffling , t dna binary system , recombinant dna , dna , gene , vector (molecular biology) , directed evolution , computer science , mutant , programming language
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 I intron ribozyme from Candida albicans . This plasmid expresses functional rRNA and ribozyme. Copyright © 2000 John Wiley & Sons, Ltd.