Cloning of the HIS5 gene of Saccharomyces cerevisiae by yeast transformation.
A yeast DNA fragment complementing a yeast his5 mutation was cloned on a shuttle vector, YRp7, by transformation of the yeast host with plasmid DNA prepared from a gene bank constructed in the Escherichia coli host. That the DNA fragment contains the yeast HIS5 gene was confirmed by the integration of the cloned plasmid into the his5 region of the yeast chromosome. The cloned yeast HIS5 gene weakly complemented the E. coli hisC mutation and gave rise to a slow-growing E. coli transformant without addition of histidine. From the slow-growing culture, rapidly growing variants of E. coli were easily obtained by mutations either in the bacterial chromosome or in the plasmid.
D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D010957:Plasmids; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D000637:Transaminases / Q000235:genetics*; D014170:Transformation, Genetic
D004271:DNA, Fungal; D000637:Transaminases