Extracellular expression of glucose inhibition-resistant Cellulomonas flavigena PN-120 β-glucosidase by a diploid strain of Saccharomyces cerevisiae

2013 ◽  
Vol 196 (1) ◽  
pp. 25-33 ◽  
Author(s):  
David J. Mendoza-Aguayo ◽  
Héctor M. Poggi-Varaldo ◽  
Jaime García-Mena ◽  
Ana C. Ramos-Valdivia ◽  
Luis M. Salgado ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Extracellular Expression ◽  
Diploid Strain ◽  
Cellulomonas Flavigena ◽  
Glucose Inhibition

scholarly journals Ribosomal protein L30 is dispensable in the yeast Saccharomyces cerevisiae.

1990 ◽  
Vol 10 (10) ◽  
pp. 5235-5243 ◽  
Author(s):  
D M Baronas-Lowell ◽  
J R Warner
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ribosomal Protein ◽  
Ribosomal Proteins ◽  
Wild Type ◽  
Diploid Strain ◽  
Chain Reaction ◽  
Yeast Saccharomyces Cerevisiae ◽  
Ribosomal Subunits ◽  
Strong Homology ◽  
Polymerase Chain

In the yeast Saccharomyces cerevisiae, L30 is one of many ribosomal proteins that is encoded by two functional genes. We have cloned and sequenced RPL30B, which shows strong homology to RPL30A. Use of mRNA as a template for a polymerase chain reaction demonstrated that RPL30B contains an intron in its 5' untranslated region. This intron has an unusual 5' splice site, C/GUAUGU. The genomic copies of RPL30A and RPL30B were disrupted by homologous recombination. Growth rates, primer extension, and two-dimensional ribosomal protein analyses of these disruption mutants suggested that RPL30A is responsible for the majority of L30 production. Surprisingly, meiosis of a diploid strain carrying one disrupted RPL30A and one disrupted RPL30B yielded four viable spores. Ribosomes from haploid cells carrying both disrupted genes had no detectable L30, yet such cells grew with a doubling time only 30% longer than that of wild-type cells. Furthermore, depletion of L30 did not alter the ratio of 60S to 40S ribosomal subunits, suggesting that there is no serious effect on the assembly of 60S subunits. Polysome profiles, however, suggest that the absence of L30 leads to the formation of stalled translation initiation complexes.

Ribosomal protein L30 is dispensable in the yeast Saccharomyces cerevisiae

1990 ◽  
Vol 10 (10) ◽  
pp. 5235-5243
Author(s):  
D M Baronas-Lowell ◽  
J R Warner
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ribosomal Protein ◽  
Ribosomal Proteins ◽  
Wild Type ◽  
Diploid Strain ◽  
Chain Reaction ◽  
Yeast Saccharomyces Cerevisiae ◽  
Ribosomal Subunits ◽  
Strong Homology ◽  
Polymerase Chain

In the yeast Saccharomyces cerevisiae, L30 is one of many ribosomal proteins that is encoded by two functional genes. We have cloned and sequenced RPL30B, which shows strong homology to RPL30A. Use of mRNA as a template for a polymerase chain reaction demonstrated that RPL30B contains an intron in its 5' untranslated region. This intron has an unusual 5' splice site, C/GUAUGU. The genomic copies of RPL30A and RPL30B were disrupted by homologous recombination. Growth rates, primer extension, and two-dimensional ribosomal protein analyses of these disruption mutants suggested that RPL30A is responsible for the majority of L30 production. Surprisingly, meiosis of a diploid strain carrying one disrupted RPL30A and one disrupted RPL30B yielded four viable spores. Ribosomes from haploid cells carrying both disrupted genes had no detectable L30, yet such cells grew with a doubling time only 30% longer than that of wild-type cells. Furthermore, depletion of L30 did not alter the ratio of 60S to 40S ribosomal subunits, suggesting that there is no serious effect on the assembly of 60S subunits. Polysome profiles, however, suggest that the absence of L30 leads to the formation of stalled translation initiation complexes.

Expression of a codon-optimized β-glucosidase from Cellulomonas flavigena PR-22 in Saccharomyces cerevisiae for bioethanol production from cellobiose

2017 ◽  
Vol 199 (4) ◽  
pp. 605-611 ◽  
Author(s):  
Francisco Javier Ríos-Fránquez ◽  
Enrique González-Bautista ◽  
Teresa Ponce-Noyola ◽  
Ana Carmela Ramos-Valdivia ◽  
Héctor Mario Poggi-Varaldo ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Bioethanol Production ◽  
Cellulomonas Flavigena

scholarly journals Disruption of single-copy genes encoding acidic ribosomal proteins in Saccharomyces cerevisiae.

1990 ◽  
Vol 10 (5) ◽  
pp. 2182-2190 ◽  
Author(s):  
M Remacha ◽  
C Santos ◽  
J P Ballesta
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ribosomal Proteins ◽  
Minimal Medium ◽  
Single Copy ◽  
Diploid Strain ◽  
Tetrad Analysis ◽  
Genes Encoding ◽  
Acidic Proteins ◽  
Dna Fragment ◽  
His3 Gene

Using the cloned genes coding for the ribosomal acidic proteins L44 and L45, constructions were made which deleted part of the coding sequence and inserted a DNA fragment at that site carrying either the URA3 or HIS3 gene. By gene disruption techniques with linearized DNA from these constructions, strains of Saccharomyces cerevisiae were obtained which lacked a functional gene for either protein L44 or protein L45. The disrupted genes in the transformants were characterized by Southern blots. The absence of the proteins was verified by electrofocusing and immunological techniques, but a compensating increase of the other acidic ribosomal proteins was not detected. The mutant lacking L44 grew at a rate identical to the parental strain in complex as well as in minimal medium. The L45-disrupted strain also grew well in both media but at a slower rate than the parental culture. A diploid strain was obtained by crossing both transformants, and by tetrad analysis it was shown that the double transformant lacking both genes is not viable. These results indicated that proteins L44 and L45 are independently dispensable for cell growth and that the ribosome is functional in the absence of either of them.

scholarly journals Whole-Genome Sequence of an Isogenic Haploid Strain, Saccharomyces cerevisiae IR-2idA30(MATa), Established from the Industrial Diploid Strain IR-2

2019 ◽  
Vol 8 (17) ◽  
Author(s):  
Kazuhiro E. Fujimori ◽  
Yosuke Kobayashi ◽  
Taisuke Seike ◽  
Takehiko Sahara ◽  
Satoru Ohgiya ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Genome Sequence ◽  
Draft Genome ◽  
Whole Genome Sequence ◽  
Whole Genome ◽  
Diploid Strain ◽  
Haploid Strain ◽  
Content Type ◽  
Sequencing Errors ◽  
Long Reads

We present the draft genome sequence of an isogenic haploid strain, IR-2idA30(MAT a), established from Saccharomyces cerevisiae IR-2. Assembly of long reads and previously obtained contigs from the genome of diploid IR-2 resulted in 50 contigs, and the variations and sequencing errors were corrected by short reads.

Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate as a rapid mapping method in Saccharomyces cerevisiae

1982 ◽  
Vol 2 (9) ◽  
pp. 1080-1087
Author(s):  
J S Wood
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mitotic Chromosome ◽  
Chromosomal Location ◽  
Mapping Method ◽  
Chromosome Loss ◽  
Diploid Strain ◽  
Simple Method ◽  
Rapid Mapping

Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate has been utilized as a rapid and simple method for assigning genes to individual chromosomes in Saccharomyces cerevisiae. This technique relied on the segregation of heterozygous markers in a diploid strain after methyl benzimidazole-2-yl-carbamate treatment due to loss of whole chromosomes. Correlations between the expression of an unmapped gene and that of a previously mapped recessive marker indicated chromosomal linkage. Depending on whether the unmapped gene and the marker were located in coupling or in repulsion, either positive or negative correlations were seen. The chromosomal location of several previously mapped genes were confirmed as a test of the method, and one previously unmapped gene, nib1, was mapped.

Disruption of single-copy genes encoding acidic ribosomal proteins in Saccharomyces cerevisiae

1990 ◽  
Vol 10 (5) ◽  
pp. 2182-2190
Author(s):  
M Remacha ◽  
C Santos ◽  
J P Ballesta
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Ribosomal Proteins ◽  
Minimal Medium ◽  
Single Copy ◽  
Diploid Strain ◽  
Tetrad Analysis ◽  
Genes Encoding ◽  
Acidic Proteins ◽  
Dna Fragment ◽  
His3 Gene

Using the cloned genes coding for the ribosomal acidic proteins L44 and L45, constructions were made which deleted part of the coding sequence and inserted a DNA fragment at that site carrying either the URA3 or HIS3 gene. By gene disruption techniques with linearized DNA from these constructions, strains of Saccharomyces cerevisiae were obtained which lacked a functional gene for either protein L44 or protein L45. The disrupted genes in the transformants were characterized by Southern blots. The absence of the proteins was verified by electrofocusing and immunological techniques, but a compensating increase of the other acidic ribosomal proteins was not detected. The mutant lacking L44 grew at a rate identical to the parental strain in complex as well as in minimal medium. The L45-disrupted strain also grew well in both media but at a slower rate than the parental culture. A diploid strain was obtained by crossing both transformants, and by tetrad analysis it was shown that the double transformant lacking both genes is not viable. These results indicated that proteins L44 and L45 are independently dispensable for cell growth and that the ribosome is functional in the absence of either of them.

Extracellular expression of the HT1 neurotoxin from the Australian paralysis tick in two Saccharomyces cerevisiae strains

Toxicon ◽  
2017 ◽  
Vol 140 ◽  
pp. 1-10 ◽  
Author(s):  
Thomas Karbanowicz ◽  
Eric Dover ◽  
Xinyi Mu ◽  
Ala Tabor ◽  
Manuel Rodriguez-Valle
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Extracellular Expression ◽  
Paralysis Tick

scholarly journals Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate as a rapid mapping method in Saccharomyces cerevisiae.

1982 ◽  
Vol 2 (9) ◽  
pp. 1080-1087 ◽  
Author(s):  
J S Wood
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Mitotic Chromosome ◽  
Chromosomal Location ◽  
Mapping Method ◽  
Chromosome Loss ◽  
Diploid Strain ◽  
Simple Method ◽  
Rapid Mapping

Mitotic chromosome loss induced by methyl benzimidazole-2-yl-carbamate has been utilized as a rapid and simple method for assigning genes to individual chromosomes in Saccharomyces cerevisiae. This technique relied on the segregation of heterozygous markers in a diploid strain after methyl benzimidazole-2-yl-carbamate treatment due to loss of whole chromosomes. Correlations between the expression of an unmapped gene and that of a previously mapped recessive marker indicated chromosomal linkage. Depending on whether the unmapped gene and the marker were located in coupling or in repulsion, either positive or negative correlations were seen. The chromosomal location of several previously mapped genes were confirmed as a test of the method, and one previously unmapped gene, nib1, was mapped.

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