Construction and restriction endonuclease mapping of hybrid plasmids containing Saccharomyces cerevisiae ribosomal DNA

Abstract

Fragments produced by partial digestion of Saccharomyces cerevisiae ribosomal DNA (rDNA) with the restriction endonuclease EcoRI were ligated in vitro to the bacterial plasmid RSF2124. The resulting hybrid plasmids were cloned in Escherichia coli. Three hybrid plasmids which contain at least one intact repetitive unit of the multiple, tandem sequences of the yeast rDNA genes have been further characterized. These plasmids have been used to construct a map of the EcoRI, SmaI, HindII and HindIII restriction sites in the individual repetitive units of yeast rDNA.

Copy number and the stability of 2-micron circle-based artificial plasmids of Saccharomyces cerevisiae

“…DNA was digested with EcoRI, run on agarose gels, and then stained with ethidium bromide. The most prominent bands seen were those due to plasmid restriction fragments, and those due to the 2.70-, 2.23-, and 1.82-kilobase (kb)EcoRI restriction fragments of rDNA (7,19). The gels were photographed by using exposures calculated to minimize the nonlinear response of the Polaroid emulsion used.…”

Tandemly arranged variant 5S ribosomal RNA genes In the yeastSaccharomyces cerevisiae

“…The yeast rR'NA gene repeat is about 9 kb in size and has a relatively small amount of non-transcribed spacer (about 2 kb). The repeat has been mapped with restriction enzymes and the position and orientation of the rRNA transcripts within the repeat has been determined (2)(3)(4)(5)(6). Most lab strains of yeast have rRNA genes that contain seven EcoRI fragments, labeled A through G in order of size (Figure la).…”

Primary structure and transcription of an amplified genetic locus: the CUP1 locus of yeast.

“…The genetic organization of the CUP] locus resembles the organization of the rRNA genes in yeast, Drosophila, Xenopus, silkworm, and sea urchin, which are arranged in clusters of tandemly arrayed repeating units (5)(6)(7)(8)(9)(10)(11)(12). Histone genes of sea urchin, Drosophila, and chicken also show a similar arrangement in tandem arrays (13)(14)(15)(16).…”