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Nucleic Acids Research, 1985, Vol. 13, No. 23 8487-8506
© 1985

Articles

A transcription map of a yeast centromere plasmid: unexpected transcripts and altered gene expression

Gregory T. Marczynski* and Judith A. Jaehning*

Department of Biochemistry, University of Illinois Urbana, IL 61801, USA

Received July 12, 1985. Revised November 13, 1985. Accepted November 13, 1985.

YCpl9 is a yeast centromere plasmid capable of autonomous replication in both yeast and E. coll (J. Mol. Biol., 158: 157–179, 1982). It is stably maintained as a single copy in the yeast cell and is therefore a model yeast "minichromosome" and cloning vector. We have located the positions and measured the abundance of the in vivo yeast transcripts from YCpl9. Transcripts from the selectable marker genes TRP1 and URA3 were present at increased levels relative to chromosomal copies of the genes. Unanticipated transcripts from the yeast CEN4 and E. coli pBR322 sequences were also found. Although much of the plasmid vector is actively transcribed in vivo, the regions around the most useful cloning sites (BamHI, EcoRI, SalI) are free of transcripts. We have analyzed transcription of BamHI inserts containing promoter variants of the HIS3 gene and determined that although initiation events are accurate, plasmid context may alter levels of gene expression.

*Present address: Department of Biology, Indiana University, Bloomington, IN 47405. USA


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