• Journal of Life Science
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• v.26 no.6
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• pp.646-650
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• 2016

The influence of chromosome number on cell growth and cell aging was investigated in various yeast strains that have many artificial chromosomes constructed using a chromosome manipulation technique. Host strain FY833 and the YKY18, YKY18R, YKY24, and YKY30 strains harboring 16 natural chromosomes, 18 chromosomes, 18 chromosomes containing rDNA chromosome, 24 chromosomes, and 30 chromosomes, respectively, were used, and the specific growth rate of each strain was compared. The specific growth rates in the YKY18 and YKY24 strains were indistinguishable from that in the host strain, while those of the YKY18R and YKY30 strains were reduced to approximately 25% and 40% of the host strain level, respectively. Subsequently, the replicative life span was examined to investigate the relationship between the number of chromosomes and cell aging, and the life span was decreased to approximately 14% and 45% of the host strain level in the YKY24 and YKY30 strains, respectively. Moreover, telomere length, well known as a senescence factor, was shorter and more diversified in the strain, showing decreased life span. Therefore, these results suggest the possibility that an increase in the number of chromosomes containing artificial chromosomes caused cell aging, and we expected these observations would be applied to improve industrial strain harboring of versatile and special artificial chromosomes.

• Journal of Life Science
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• v.21 no.9
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• pp.1226-1233
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• 2011

In neurons, kinesin is the molecular motor that transport cargos along microtubules. KIF5s (alias kinesin-I), are heterotetrameric motor conveying cargos, but the mechanism as to how they recognize and bind to a specific cargos has not yet been completely elucidated. To identify the interaction proteins for KIF5C, yeast two-hybrid screening was performed, and specific interaction with the $\underline{S}$am68-$\underline{l}$ike $\underline{m}$ammalian protein $\underline{2}$ (SLM-2), a member of the $\underline{s}$ignal $\underline{t}$ransducers and $\underline{a}$ctivators of $\underline{R}$NA (STAR) family of RNA processing proteins, was found. SLM-2 bound to the carboxyl (C)-terminal region of KIF5C and to other KIF5 members. The C-terminal domain of Sam68, SLM-1, SLM-2 was essential for interaction with KIF5C in the yeast two-hybrid assay. In addition, glutathione S-transferase (GST) pull-downs showed that SAM68, SLM-1, and SLM-2 specifically interacted to Kinesin-I complex. An antibody to SAM68 specifically co-immunoprecipitated SAM68 associated with KIF5s and coprecipitated with a specific set of mRNA. These results suggest that Kinesin-I motor protein transports RNA-associated protein complex in cells.

• The Korean Journal of Mycology
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• v.43 no.1
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• pp.47-52
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• 2015

Several kinds of yeasts were isolated from wild flowers collected from Muju resort of Deogyu mountain in Korea and identified by comparison of nucleotide sequences of D1/D2 region of 26S rDNA using BLAST. Totally twenty-two strains of eight species were isolated and Cryptococcus magnus (9 strains) and Cryptococcus flavescenes (6 strains) were dominant species. The supernatant and cell-free extracts were prepared and their antioxidant activity, anti-gout xanthine oxidase inhibitory activity and whitening tyrosinase inhibitory activity were investigated. Anti-gout xanthine oxidase inhibitory activities of Sporobolomyces phaffii JS00583 and Rhodotorula graminis JS00581 were 40.4% and 39.1%, respectively and also was 36.1% whitening tyrosinase inhibitory activity in the supernatant of Cryptococcus magnus JS00570.

• KSBB Journal
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• v.11 no.3
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• pp.270-275
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• 1996

Heterologous expression of glucose oxidase gene using recombinant yeast has been carried out. Polymerase chain reaction was conducted to obtain the gene encoding glucose oxidase from Aspergillus niger and sequence comparison indicated the cloned 1.9kb DNA fragment appeared to be the glucose oxidise structural gene containing a signal sequence for extracellular location. Transforming shuttle vector was constructed with YEp352 to express the cloned glucose oxidase gene under the control of either GAL1 or GAL10 promoter. Plate assay of recombinant yeasts has shown that GAL1 promoter was more effective in yielding glucose oxidise than GAL10 promoter. Among the five different concentrations of galactose tried, 1% galactose showed the highest induction of glucose oxidase. Cellular localization experiment of recombinant enzyme using spheroplast revealed that most of enzymes (80%) were secreted into culture media in contrast to A. niger. There is no difference in heat-stability of recombinant enzyme up to $50^{\circ}C$ compared to the glucose oxidase from A. niger However, a dramatic reduction of enzyme activity was observed in both enzymes at $60^{\circ}C$.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.120-127
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• 1990

Intraspecific fusion frequency between Filobasidium capsuligenum CBS6122-2 ade and met trp or met strains was $2.9\times 10^{-3}$ and $8.5\times 10^{-3}$, respectively. And intergeneric fusion frequency between Folobadidium capsuligenum CBS 4318 (cys his) and Saccharomyces cerevisiae 262-12-1 (hom3 thr1) or X2180-1A (ade thr) was $8.8\times 10^{-6}$ and $9.5\times 10^{-4}$, respectively. Nuclear fusion appears to occru in fusants between intraspecies and intergenera as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and mitotic segregation analysis. It was also found that $\alpha$-amylase and glucoamylase activity from intraspecific hybrids was 1.6-2.1 fold increased when compared with that from thier parents.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.249-253
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• 2014

THOC7/Mft1 is a member of the THO complex that is an evolutionally conserved assembly connecting transcription elongation with mRNP packaging and mRNA export. In fission yeast Schizosaccharomyces pombe, an ortholog (spThoc7) of THOC7/Mft1 was isolated by partial complementation of the lethality in synthetic lethal mutant, SLRsm1. A deletion mutant in a diploid strain was constructed by replacing one of spThoc7-coding region with an $ura4^+$ gene using one-step gene disruption method. Tetrad analysis showed that the spthoc7 is nonessential for growth. But the ${\Delta}thoc7$ null mutant showed slight defects of both growth and mRNA export. And the functional spThoc7-GFP protein is localized mainly in the nucleus. These results suggest that spThoc7 is also involved in mRNA export from the nucleus to cytoplasm.

• Korean Journal of Microbiology
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• v.51 no.2
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• pp.181-185
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• 2015

Tho2/THOC2 is a subunit of the THO complex that plays important roles in mRNP biogenesis connecting transcription with mRNA maturation and export. A fission yeast, Schizosaccharomyces pombe, ortholog of Tho2/THOC2 was identified from the genome database. Tetrad analysis showed that the S. pombe tho2 is essential for growth. Repression or overexpression of the tho2 gene caused growth defect with elongated cells, abnormal DNA distribution, and accumulation of $poly(A)^+$ RNA in the nucleus. And the functional GFP-Tho2 protein is localized mainly in the nucleus. Yeast two-hybrid analysis showed that Tho2 interacted with Tex1, another subunit of THO complex. These results suggest that S. pombe Tho2 is also involved in mRNA export from the nucleus and is a component of THO complex.

• Journal of Life Science
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• v.17 no.1 s.81
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• pp.39-44
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• 2007

The Rqh1 gene is essential for vegetative growth in fission Yeast. The rqh1 mutant showed that sensitivity of DNA damaging agent, a wild range of phenotype including abnormal gene expression and cell elongation. This result showed that the rqhl-overexpression cell was sensitivity to DNA damaging agent like rqhl mutant. When Rqh1 have an over-expression by $nmt1^+$ promoter of pREP vector, rqh1 mutant DNA damaging agent sensitivity could be compensated. We isolated two strong mutant containing complementation gene, rqh156 and rqh172, respectively. This result observed that the DNA damaging agent sensitivity of rqhl mutant was complemented by the expression of rqh156 and rqh172. They induced mRNA expression in a dose-dependent manner HU, MMS and UV. The HU sensitivity of the rqhl was complemented by the expression of rqh156 and rqh172. The mRNA expression of rqh156 decreased on HU dose dependent but the mRNA expression of rqh172 did not decrease on HU dose dependent. The MMS and W sensitivity of the rqhl was complemented by the expression of rqh156 and rqh172. These results indicate that the isolated rqhl gene may play an important role in DNA metabolism.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.116-119
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• 2016

Stable maintenance of telomere is required for cell proliferation and survival. Although telomerase is the primary means for telomere maintenance, recombination is another important pathway to maintain telomeres. In this study, we developed a genetic assay for telomere recombination using the internal $TG_{1-3}$ repeats present in subtelomeric regions of yeast. The recombination frequencies were dependent on the presence of the internal $TG_{1-3}$ repeats. PCR amplification of the regions near URA3 and CAN1 markers using genomic DNA isolated from $FOA^rCan^r$ colonies indicated that each isolate had lost the chromosome end including the markers. In addition, the recombination frequencies increased with longer internal $TG_{1-3}$ repeats. Our results suggest that the $FOA^rCan^r$ colony formation is the consequence of recombination between the internal and terminal $TG_{1-3}$ repeats.

• Korean Journal of Microbiology
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• v.53 no.4
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• pp.235-241
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• 2017

The purpose of this study was to investigate the enhanced physiological activities of two Pichia strains, yeasts isolated from grapes pericarp. Based on phylogenetic analysis using 18S rRNA sequencing, two isolates were identified as Pichia manshurica GU-3 and Pichia terricola GU-4, respectively. The scanning electron microscopic analysis showed that the two isolates grown on YPD medium were of typical elliptical shape with buds and bud scars on cell surface. Physiological activities of the single and mixed Pichia cultures were monitored and compared. In mixed cultures after 72 h of incubation, the maximum activities of tyrosinase inhibition, ACE inhibition, and antioxidant were 81.7%, 45.9%, and 42.7%, respectively. Superoxide dismutase-like activity was approximately 30% in the mixed cultures. These studies demonstrate that Pichia species cultured in the form of mixture can enhance the physiological activities and has potential for the development of new bioactive products.