Chromosomal DNA preparation from yeasts of biotechnological importance (original) (raw)
Applied and Environmental Microbiology, 2009
Yeasts used in the production of lagers contain complex allopolyploid genomes, resulting from the fusion of two different yeast species closely related to Saccharomyces cerevisiae and Saccharomyces bayanus. Recombination between the homoeologous chromosomes has generated a number of hybrid chromosomes. These recombination events provide potential for adaptive evolution through the loss or gain of gene function. We have examined the genotypic and phenotypic effects of one of the conserved recombination events that occurred on chromosome XVI in the region of YPR159W and YPR160W. Our analysis shows that the recombination event occurred within the YPR160W gene, which encodes the enzyme glycogen phosphorylase and generates a hybrid gene that does not produce mature mRNA and is nonfunctional due to frameshifts in the coding region. The loss of function of the hybrid gene leads to glycogen levels similar to those found in haploid yeast strains. The implications for the control of glycogen levels in fermentative yeasts are discussed.
Yeast Genetics and Biotechnological Applications
Yeast Biotechnology: Diversity and Applications, 2009
Contents for conducting classical genetics. Various vector systems available, foreign genes expressed and the limitations as a host will be discussed. Finally, the use of various yeast enzymes in biotechnology sector will be reviewed.
The ploidy determination of the biotechnologically important yeast Candida utilis
Journal of Applied Genetics, 2020
Yeast Candida utilis is considered to be a potentially advantageous expression system for production of recombinant proteins utilizable for industrial and pharmaceutical purposes. As the scientific literature is not consistent in the ploidy of this yeast, in this work, we focused on resolving the problem via several methods such as the copy number determination of maltase gene by multiplex PCR, measuring α-glucosidase activity, the characterization of maltase gene copy number in deletion mutants using qPCR and flow cytometry. In context with the published data and results obtained in this study about the copy number of the maltase gene on C. utilis genome, we attempted to hypothesise and made conclusion about the ploidy of C. utilis. The results of this work, besides the biotechnological aspect, contribute to the elementary knowledge of C. utilis. The exact information about the ploidy or more specifically about the copy number of appropriate gene is essential for expression cassette dosage determination integrated into the chromosome of the host. In this study, we come to the conclusion that the maltase gene is present in C. utilis genome in four alleles, and in combination with flow cytometry, published information and the published genome sequences, the observations support the theory about tetraploidy of C. utilis.
Characterization of two types of yeast ribosomal DNA genes
Journal of bacteriology, 1978
The intragenic organization of ribosomal DNA from a diploid strain of Saccharomyces cerevisiae was analyzed by using recombinant DNA molecules constructed in vitro. Restriction analysis of the yeast ribosomal DNA with the EcoRI restriction enzyme indicated that eight restriction fragments were present in the ribosomal DNA of this strain: X' (1.87 X 10(6) daltons), A (1.77 X 10(6) daltons), B (1.48 X 10(6) daltons), C (1.22 X 10(6) daltons), D (0.39 X 10(6) daltons), E (0.36 X 10(6) daltons), F (0.22 X 10(6) daltons), and G (0.17 X 10(6) daltons). These fragments were distributed between two different types of ribosomal DNA genes, which had the restriction maps: (formula: see text) in which the underlined region shows the repeating unit. The diploid yeast strain contained approximately equal amounts of each of these two types of genes. The analysis of the recombinant DNA molecules also indicated that the yeast ribosomal genes are homogeneous and extensively clustered.
Brazilian Archives of Biology and Technology, 2012
The purpose of this work was to study a rapid yeast DNA extraction by boiling and freeze-thawing processes without using chemical reagents or any purification procedures, to obtain a high grade PCR-product. A specific DNA fragment of the 18S region of Dekkera bruxellensis and Saccharomyces cerevisiae was chosen. The described boiling and freeze-thawing protocols generated the PCR-grade product preparations and could be used to process many samples. The amplification of the fragments could be observed after 30 and 35 cycles. These processes of extraction without using any kind of chemical reagents, especial water, and purification procedures proved to be efficient, reproducible, simple, fast, and inexpensive.
Regeneration of yeast protoplasts in agar gels
Experimental Cell Research, 1966
FmM human lymphocyte nuclei we have extracted DNA fibres with lengths ranging up to 2.2 cm. Such long fibres are to be expected if the interphase chromosome consists basically of a single DNA fibre.
Nucleic acid reduction in yeast
Applied Microbiology and Biotechnology, 1988
A method for reduction of nucleic acid levels in preparations of the yeasts Saccharomyces cerevisiae and Kluyveromycesfragilis by means of alkaline treatment has been developed. Under similar conditions (4.5% NH4OH, 65 ° C, 30 min) a low nucleic acid content (less than 2%) was obtained for both strains. Higher losses of proteins and biomass were obtained with K. fragilis than with S. cerevisiae. 1977). The purpose of the present work was to obtain the reduction of nucleic acid levels in two yeast strains by a moderate treatment with NHaOH. Saccharomyces cerevisiae (baker's yeast), widely used in several foods, and Kluyveromyces fragilis, proposed as one of the more suitable yeasts for human consumption (Contreras and Vald6s 1984), were employed.
Isolation of an episomal yeast gene and replication origin as chromatin
Proceedings of the National Academy of Sciences, 1986
A multicopy yeast plasmid containing the TRPI gene (coding for N-5'-phosphoribosylanthranilate isomerase) and ARSI (autonomously replicating sequence 1) has been purified as chromatin. Electrophoretic analysis of nucleic acid and proteins and electron microscopy show that the plasmid chromatin is largely free of contaminants. Electronmicroscopic and linking-number analyses indicate that the plasmid chromatin contains seven nucleosomes, as predicted by the indirect end-label analyses of Thoma, Bergman, and Simpson U. Mol. Biol. (1984) 177, 715-733]. Indirect end label
Comercial yeast samples DNA extraction using universal kit
2021
Yeast is a microorganism that is relatively difficult to obtain its DNA mass. This is caused by the content of cell walls with a thick and rigid composition, such as chitin. This study aims to provide information on the best concentration and purity of the DNA extraction results of various types of commercial yeast in different samples weight. The method used is based on Universal Kit procedure with modification. This study indicates that differences in concentration or sample weight do not affect the DNA concentration and purity ratio value of each yeast sample. Each best result obtained from Jago (Rhizopus oligosporus), Raprima (Rhizopus oligosporus) and Fermipan (Saccharomyces cerevisiae) in 50 mg sample, NKL (Saccharomyces cerevisiae) yeast in 40 mg. Recomendation weight samples to extract DNA of Rhizopus oligosporus and Saccharomyces cerevisiae isolates are between 40 mg to 50 mg. The contamination factor causes the difference in the value of purity in each sample during DNA extraction.
Revista de Microbiologia, 1999
In order to characterize fusion products from yeast protoplasts and their segregants, with important features to the wine making industry, electrophoretic karyotyping and RAPD (Random Amplified Polymorphic DNA) were utilized. Electrophoretic karyotyping was performed by the CHEF ("contour-clamped homogeneous electric field electrophoresis") method, which allowed the detection of chromosomal band complementation in fusion products and the presence of patterns of both parental and intermediary strains in segregants. By utilizing two primers, an amplification pattern of DNA fragments was obtained. While fusion products (diploid) showed a pattern of complementary bands, segregants showed bands of either parental strains or even intermediary bands