scholarly journals Molecular cloning and characterization of the threonine deaminase (ILV1) gene of Saccharomyces cerevisiae

1983 ◽  
Vol 48 (3) ◽  
pp. 149-159 ◽  
Author(s):  
Jens G. Litske Petersen ◽  
Steen Holmberg ◽  
Torsten Nilsson-Tillgren ◽  
Morten C. Kielland-Brandt
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Cloning ◽  
Threonine Deaminase

Molecular cloning and characterization of the RAD1 gene of Saccharomyces cerevisiae

Gene ◽  
1983 ◽  
Vol 26 (2-3) ◽  
pp. 119-126 ◽  
Author(s):  
David R. Higgins ◽  
Satya Prakash ◽  
Paul Reynolds ◽  
Louise Prakash
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Cloning

Molecular cloning of eucaryotic genes required for excision repair of UV-irradiated DNA: isolation and partial characterization of the RAD3 gene of Saccharomyces cerevisiae

1982 ◽  
Vol 152 (1) ◽  
pp. 323-331
Author(s):  
L Naumovski ◽  
E C Friedberg
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Cloning ◽  
Dna Isolation ◽  
Excision Repair ◽  
Restriction Analysis ◽  
Uv Resistance ◽  
Chromosomal Dna ◽  
Uv Sensitivity ◽  
Resultant Plasmid

We describe the molecular cloning of a 6-kilobase (kb) fragment of yeast chromosomal DNA containing the RAD3 gene of Saccharomyces cerevisiae. When present in the autonomously replicating yeast cloning vector YEp24, this fragment transformed two different UV-sensitive, excision repair-defective rad3 mutants of S. cerevisiae to UV resistance. The same result was obtained with a variety of other plasmids containing a 4.5-kb subclone of the 6-kb fragment. The UV sensitivity of mutants defective in the RAD1, RAD2, RAD4, and RAD14 loci was not affected by transformation with these plasmids. The 4.5-kb fragment was subcloned into the integrating yeast vector YIp5, and the resultant plasmid was used to transform the rad3-1 mutant to UV resistance. Both genetic and physical studies showed that this plasmid integrated by homologous recombination into the rad3 site uniquely. We conclude from these studies that the cloned DNA that transforms the rad3-1 mutant to UV resistance contains the yeast chromosomal RAD3 gene. The 4.5-kb fragment was mapped by restriction analysis, and studies on some of the subclones generated from this fragment indicate that the RAD3 gene is at least 1.5 kb in size.

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