Mutations at the Saccharomyces cerevisiae SUP4 tRNA Tyr Locus: Isolation, Genetic Fine-Structure Mapping, and Correlation with Physical Structure

1982 ◽  
Vol 2 (12) ◽  
pp. 1501-1513
Author(s):  
Janet Kurjan ◽  
Benjamin D. Hall

The SUP4 tRNA Tyr locus in Saccharomyces cerevisiae has been studied by the isolation and characterization of mutations at the SUP4 gene which result in the loss of suppressor function. Most of the mutations act as single-site mutations, whereas about a third of the mutations are deletions of the entire gene. Two meiotic fine-structure maps of the gene were made. The first mapping technique placed 10 mutations plus the sup4 + anticodon on a map by a measurement of levels of recombination between pairs of mutations. The second map utilized a more qualitative estimate of recombination frequency, allowing 69 mutations and the sup4 + anticodon to be mapped. The maps were compared with the physical structure of the gene for the 34 mutations whose nucleotide alteration has been determined by DNA sequencing (Koski et al., Cell 22 :415-425, 1980; Kurjan et al., Cell 20 :701-709, 1980). Both maps show a good correlation with the physical structure of the gene, even though certain properties of genetic fine-structure maps, such as marker effects and “map expansion,” were seen.

1982 ◽  
Vol 2 (12) ◽  
pp. 1501-1513 ◽  
Author(s):  
Janet Kurjan ◽  
Benjamin D. Hall

TheSUP4tRNATyrlocus in Saccharomyces cerevisiae has been studied by the isolation and characterization of mutations at theSUP4gene which result in the loss of suppressor function. Most of the mutations act as single-site mutations, whereas about a third of the mutations are deletions of the entire gene. Two meiotic fine-structure maps of the gene were made. The first mapping technique placed 10 mutations plus thesup4+anticodon on a map by a measurement of levels of recombination between pairs of mutations. The second map utilized a more qualitative estimate of recombination frequency, allowing 69 mutations and thesup4+anticodon to be mapped. The maps were compared with the physical structure of the gene for the 34 mutations whose nucleotide alteration has been determined by DNA sequencing (Koski et al., Cell22:415-425, 1980; Kurjan et al., Cell20:701-709, 1980). Both maps show a good correlation with the physical structure of the gene, even though certain properties of genetic fine-structure maps, such as marker effects and “map expansion,” were seen.


1984 ◽  
Vol 4 (1) ◽  
pp. 86-91 ◽  
Author(s):  
G T Maine ◽  
R T Surosky ◽  
B K Tye

We have cloned a functional centromeric DNA sequence from Saccharomyces cerevisiae. Using the 2 mu chromosome-loss mapping technique and meiotic tetrad analysis, we have identified this DNA sequence as the centromere of chromosome V (CEN5). The CEN5 sequence has been localized on an 1,100-base-pair BamHI-BglII restriction fragment. Plasmids containing CEN5 and an autonomously replicating sequence are mitotically stable in S. cerevisiae and segregate in a Mendelian fashion during meiosis.


1984 ◽  
Vol 4 (1) ◽  
pp. 86-91
Author(s):  
G T Maine ◽  
R T Surosky ◽  
B K Tye

We have cloned a functional centromeric DNA sequence from Saccharomyces cerevisiae. Using the 2 mu chromosome-loss mapping technique and meiotic tetrad analysis, we have identified this DNA sequence as the centromere of chromosome V (CEN5). The CEN5 sequence has been localized on an 1,100-base-pair BamHI-BglII restriction fragment. Plasmids containing CEN5 and an autonomously replicating sequence are mitotically stable in S. cerevisiae and segregate in a Mendelian fashion during meiosis.


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