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

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

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 journal › Article › peer-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 language	English (US)
Pages (from-to)	1527-1534
Number of pages	8
Journal	Yeast
Volume	16
Issue number	16
DOIs	https://doi.org/10.1002/1097-0061(200012)16:16<1527::AID-YEA643>3.0.CO;2-8
State	Published - 2000
Externally published	Yes

ASJC Scopus subject areas

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

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AU - Leibowitz, Michael J

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AB - 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.

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ER -

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

Abstract

ASJC Scopus subject areas

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