Cloning, sequencing and disruption of the ARG8 gene encoding acetylornithine aminotransferase in the petite‐negative yeast Kluyveromyces lactis

Janssen, Antje; Chen, Xin Jie

doi:10.1002/(sici)1097-0061(199802)14:3<281::aid-yea212>3.3.co;2-5

Cited by 4 publications

(4 citation statements)

References 9 publications

(12 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It was closer to the Schizosaccharomyces pombe protein than to the S. cerevisiae one, Arg8p (encoded by YOL140w). Moreover, this sequence did not correspond to the K. lactis ARG8 gene product which has been cloned and whose disruption led to an arginine auxotrophic phenotype [28]. Analysis of the K. lactis genes which belong to this second category showed that the di¡erence between K. lactis and S. cerevisiae is not the presence/absence of orthologous genes but rather the presence/absence and the composition of gene families.…”

Section: Comparison With Other Organismsmentioning

confidence: 99%

Genomic Exploration of the Hemiascomycetous Yeasts: 11. Kluyveromyces lactis

et al. 2000

View full text Add to dashboard Cite

Random sequencing of the Kluyveromyces lactis genome allowed the identification of 2235^2601 open reading frames (ORFs) homologous to S. cerevisiae ORFs, 51 ORFs which were homologous to genes from other species, 64 tRNAs, the complete rDNA repeat, and a few Ty1-and Ty2-like sequences. In addition, the complete sequence of plasmid pKD1 and a large coverage of the mitochondrial genome were obtained. The global distribution into general functional categories found in Saccharomyces cerevisiae and as defined by MIPS is well conserved in K. lactis. However, detailed examination of certain subcategories revealed a small excess of genes involved in amino acid metabolism in K. lactis. The sequences are deposited at EMBL under the accession numbers AL424881Â L430960. ß 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

show abstract

Section: Comparison With Other Organismsmentioning

confidence: 99%

Genomic Exploration of the Hemiascomycetous Yeasts: 11. Kluyveromyces lactis

et al. 2000

View full text Add to dashboard Cite

show abstract

“…S. cerevisiae genes were cloned by the complementation of S. cerevisiae mutants with S. cerevisiae plasmid libraries (Beggs, 1978;Hinnen et al, 1978;Hsiao and Carbon, 1979) and later genomic DNA libraries consisting of DNA fragments randomly inserted into yeast vectors using E. coli as a cloning host have been used for complementation cloning (Lundblad, 2001;Rose and Broach, 1991). Therefore, the cloning of genes into plasmid vectors was a common tool for gene manipulations in yeast as well as other species (Bergkamp et al, 1993;Janssen and Chen, 1998). In the present study, we demonstrate that the direct transformation of unmodified PCRamplified DNA is highly effective for the identification of auxotrophic genes in K. marxianus.…”

Section: Identification Of K Marxianus Auxotrophic Mutants With Genementioning

confidence: 99%

Identification of auxotrophic mutants of the yeast Kluyveromyces marxianus by non‐homologous end joining‐mediated integrative transformation with genes from Saccharomyces cerevisiae

Yarimizu

Nonklang

Nakamura

et al. 2013

Yeast

View full text Add to dashboard Cite

The isolation and application of auxotrophic mutants for gene manipulations, such as genetic transformation, mating selection and tetrad analysis, form the basis of yeast genetics. For the development of these genetic methods in the thermotolerant fermentative yeast Kluyveromyces marxianus, we isolated a series of auxotrophic mutants with defects in amino acid or nucleic acid metabolism. To identify the mutated genes, linear DNA fragments of nutrient biosynthetic pathway genes were amplified from Saccharomyces cerevisiae chromosomal DNA and used to directly transform the K. marxianus auxotrophic mutants by random integration into chromosomes through non-homologous end joining (NHEJ). The appearance of transformant colonies indicated that the specific S. cerevisiae gene complemented the K. marxianus mutant. Using this interspecific complementation approach with linear PCR-amplified DNA, we identified auxotrophic mutations of ADE2, ADE5,7, ADE6, HIS2, HIS3, HIS4, HIS5, HIS6, HIS7, LYS1, LYS2, LYS4, LYS9, LEU1, LEU2, MET2, MET6, MET17, TRP3, TRP4 and TRP5 without the labour-intensive requirement of plasmid construction. Mating, sporulation and tetrad analysis techniques for K. marxianus were also established. With the identified auxotrophic mutant strains and S. cerevisiae genes as selective markers, NHEJ-mediated integrative transformation with PCR-amplified DNA is an attractive system for facilitating genetic analyses in the yeast K. marxianus.

show abstract

“…cerevisiae/E. coli shuttle vector pCXJ22 (Janssen and Chen 1998). Approximately 30 000 Ura + transformants were replica-plated on GlyYP and 10 respiration competent (Gly + ) colonies were scored after incubation at 30°C for 2 days.…”

Section: Isolation Of Klatp5mentioning

confidence: 99%

Suppression of ρ 0 lethality by mitochondrial ATP synthase F1 mutations in Kluyveromyces lactis occurs in the absence of F0

1998

View full text Add to dashboard Cite

Specific mgi mutations in the alpha, beta or gamma subunits of the mitochondrial F1-ATPase have previously been found to suppress rho0 lethality in the petite-negative yeast Kluyveromyces lactis. To determine whether the suppressive activity of the altered F1 is dependent on the F0 sector of ATP synthase, we isolated and disrupted the genes KlATP4, 5 and 7, the three nuclear genes encoding subunits b, OSCP and d. Strains disrupted for any one, or all three of these genes are respiration deficient and have reduced viability. However a strain devoid of the three nuclear genes is still unable to lose mitochondrial DNA, whereas a mgi mutant with the three genes inactivated remains petite-positive. In the latter case, rho0 mutants can be isolated, upon treatment with ethidium bromide, that lack six major F0 subunits, namely the nucleus-encoded subunits b, OSCP and d, and the mitochondrially encoded Atp6, 8 and 9p. Production of rho0 mutants indicates that an F1-complex carrying a mgi mutation can assemble in the absence of F0 subunits and that suppression of rho0 lethality is an intrinsic property of the altered F1 particle.

show abstract

Cloning, sequencing and disruption of the ARG8 gene encoding acetylornithine aminotransferase in the petite‐negative yeast Kluyveromyces lactis

Cited by 4 publications

References 9 publications

Genomic Exploration of the Hemiascomycetous Yeasts: 11. Kluyveromyces lactis

Genomic Exploration of the Hemiascomycetous Yeasts: 11. Kluyveromyces lactis

Identification of auxotrophic mutants of the yeast Kluyveromyces marxianus by non‐homologous end joining‐mediated integrative transformation with genes from Saccharomyces cerevisiae

Suppression of ρ 0 lethality by mitochondrial ATP synthase F1 mutations in Kluyveromyces lactis occurs in the absence of F0

Contact Info

Product

Resources

About