• Title/Summary/Keyword: Temperature sensitive mutant

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Isolation and Analysis of Temperature Sensitive Mutants in Aspergillus nidulans (Aspergillus nidulans 온도감수성 돌연변이주의 분리 및 분석)

  • 박찬규;강현삼
    • Korean Journal of Microbiology
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    • v.16 no.3
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    • pp.111-121
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    • 1978
  • About 40 temperature-sensitive mutants have been isolated as a preliminary step to study the spore germination, the cell cycle, and the control of macromolecular synthesis in Aspergillus nidulans. To obtain temperature-sensitive mutants rapidly and effectively, the selective enrichment method using antifungal antibiotic nystatin was developed. Based on the data which had applied to the concentration of auxotrophic mutants by the earlier investigators, the optimal concentration and the time of treatment at the nonpermissive temeprature were determined as 50 to 100 units per ml and 4.5 hr., respectively. Out of 41 ts mutants assigned to the strain symbol PK, thirteen that seemed to be arrested at the earlystage of spore germination were subjected to the further cytological and genetic analysis. Elght of these mutants are able to form germ tube and five not. Staining with acid fuchsin for the 5PK strains shows that one irreversible mutant, PK6 strain able to form germ tube, accumulate mitotic spindle, being arrested in mitosis. Another PK15 and PK23 strain have more than one intact nucleolus without germ tube formation at the restrictive temperature. the temperature-senstive mutation in PK12 strain, the onlystrain which is able occurred in certain gene specific for the germination of spore. All of the ts markers are recessive and complement each other in heterokaryon between two different ts markers at the restrictive temperature.

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Brassinosteroids-mediated regulation of ABI3 is involved in high-temperature induced early flowering in plants

  • Hong, Jeongeui;Sung, Jwakyung;Ryu, Hojin
    • Journal of Plant Biotechnology
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    • v.45 no.2
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    • pp.83-89
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    • 2018
  • The interplay of plant hormones is one of the essential mechanisms for plant growth and development. A recent study reported that Brassinosteroids (BR) and ABSCISIC ACID (ABA) interact antagonistically in early seedling developments through the BR-mediated epigenetic repression of ABSCISIC ACID-INSENSITIVE 3 (ABI3). However, the other physiological roles of the BR-mediated regulation of ABI3 and ABA responses beyond early seedling developments remain largely unknown. Here, we showed that the activation of BR signaling by high temperatures promotes flowering time through the suppression of ABI3 expressions. Elevated ambient temperature induced early flowering in wild type Col-0 plants, but not in BR-defective bri1-116 mutant plants. Conversely, a hyper BR biosynthetic dwf4-D mutant displayed more sensitive thermomorphic long shoot elongation and early flowering. Both expression patterns and physiological responses supported the biological roles of ABI3 in the regulation of floral transition and reproduction under high temperature conditions. Finally, we confirmed that the lowered expressions of the transcript and protein levels of ABI3 brought on by elevated temperature were correlated with warmth-induced early flowering phenotypes. In conclusion, our data suggest that the BR- and warmth-mediated regulation of ABI3 are important in thermomorphic reproductive phase transitions in plants.

Temperature Effect on the Productivity of Recombinant Protein in a Lysis and DNA packaging-deficient and Temperature-sensitive Bacteriophage $\lambda$System (용균과 DNA 패키징 유전자가 결핍된 온도 민감성 박테리오 파아지 람다 시스템에서 재조합 단백질 생산성에 미치는 온도의 영향)

  • Oh, Jeong-Seok;Park, Tai-Hyun
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.112-115
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    • 2005
  • E. coli in combination with bacteriophage $\lambda$ was used to overcome the intrinsic plasmid instability that is frequently found in recombinant fermentation especially in long-term operation. In order to enhance the stability and productivity, the bacteriophage ${\lambda}NM1070$ was used in this study. It is a $\lambda$ mutant, which is deficient in the synthesis of protein related to DNA packaging and cell lysis. The ${\lambda}NM1070$ is also a temperature-sensitive mutant. To optimize the production of recombinant protein in this temperature-sensitive system, the temperature effects on growth and cloned gene expression were investigated for stable and efficient recombinant gene expression. The induction to the lytic state was not complete at $36^{\circ}C$ while the temperature above $40^{\circ}C$ induced the lytic state completely. However, the productivity was decreased at $42^{\circ}C$ by temperature inhibition. The L-free cell concentration increased with the increase of temperature until $40^{\circ}C$. In conclusion, ${\lambda}NM1070$ has the optimal temperature at $38^{\circ}C$ for stability and at $40^{\circ}C$ for expression.

A transcription factor "OsNAC075" is essential for salt resistance in rice (Oryza sativa L.)

  • Jung, Yu-Jin;Lee, Myung-Chul;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.94-104
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    • 2011
  • Salt stress is a major environmental factor influencing plant growth and development. To identify salt tolerance determinants, we systematically screened salt sensitive rice mutants by use of the Activator/Dissociation (Ac/Ds) transposon tagging system. In this study, we focused on the salt sensitive mutant line, designated SSM-1. A gene encoding a NAC transcription factor homologue was disrupted by the insertion of a Ds transposon into SSM-1 line. The OsNAC075 gene (EU541472) has 7 exons and encodes a protein (486-aa) containing the NAC domain in its N-terminal region. Sequence comparison showed that the OsNAC075 protein had a strikingly conserved region at the N-terminus, which is considered as the characteristic of the NAC protein family. OsNAC075 protein was orthologous to Arabidopsis thaliana ANAC075. Phylogenetic analysis confirmed OsNAC075 belonged to the OsNAC3 subfamily, which plays an important role in response to stress stimuli. RT-PCR analysis showed that the expression of OsNAC075 gene was rapidly and strongly induced by stresses such as NaCl, ABA and low temperature ($4^{\circ}C$). Our data suggest that OsNAC075 holds promising utility in improving salt tolerance in rice.

Isolation and Characterization of Salt Tolerant Mutations in Budding Yeast Saccharomyces cerevisiae

  • Kim, Yung-Jin;Seo, Soo-Boon;Park, Shi-Young
    • Journal of Life Science
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    • v.9 no.1
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    • pp.22-25
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    • 1999
  • In order to study the mechanism for the adaptation to salt stress, we mutagenized budding yeast Saccharomyces cerevisiae with Ethylmethane sulfonate, and isolated salt-tolerant mutants. Among the salt-tolerant mutants, two strains exhibit additional temperature sensitive phenotype. Here, we report that these two salt-tolerant mutants are specific to {TEX}$Na^{+}${/TEX} rather than general osmotic stress. These mutant strains may contain mutations in the genes involved in {TEX}$Na^{+}${/TEX} home-ostasis.

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Cloning of RNA1 Gene from Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 RNA1 유전자의 클로닝)

  • 송영환;고상석;이영석;강현삼
    • Korean Journal of Microbiology
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    • v.27 no.2
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    • pp.77-84
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    • 1989
  • The temperature sensitive (ts) mutation on RNA1 gene of Saccharomyces cerevisiae prevents growth at restrictive temperature ($36^{\circ}C$) by accumulation of precursor tRNA, rRNA and mRNA (Hutchison et al., 1969; Shiokawa and Pogo, 1974; Hopper et al., 1978). RNA1 gene was cloned by complementation of the temperature sensitive growth defect of an rna1-1 mutant strain and identified by retransformation and concomitant loss of recombinant plasmid on non-selective condition. By deletion mapping, it was found that RNA1 gene resides within 3.5kb of BgII fragment.

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Diversification of the molecular clockwork for tissue-specific function: insight from a novel Drosophila Clock mutant homologous to a mouse Clock allele

  • Cho, Eunjoo;Lee, Euna;Kim, Eun Young
    • BMB Reports
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    • v.49 no.11
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    • pp.587-589
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    • 2016
  • The circadian clock system enables organisms to anticipate the rhythmic environmental changes and to manifest behavior and physiology at advantageous times of the day. Transcriptional/translational feedback loop (TTFL) is the basic feature of the eukaryotic circadian clock and is based on the rhythmic association of circadian transcriptional activator and repressor. In Drosophila, repression of dCLOCK/CYCLE (dCLK/CYC) mediated transcription by PERIOD (PER) is critical for inducing circadian rhythms of gene expression. Pacemaker neurons in the brain control specific circadian behaviors upon environmental timing cues such as light and temperature cycle. We show that amino acids 657-707 of dCLK are important for the transcriptional activation and the association with PER both in vitro and in vivo. Flies expressing dCLK lacking AA657-707 in $Clk^{out}$ genetic background, homologous to the mouse Clock allele where exon 19 region is deleted, display pacemaker-neuron-dependent perturbation of the molecular clockwork. The molecular rhythms in light-cycle-sensitive pacemaker neurons such as ventral lateral neurons ($LN_vs$) were significantly disrupted, but those in temperature-cycle-sensitive pacemaker neurons such as dorsal neurons (DNs) were robust. Our results suggest that the dCLK-controlled TTFL diversify in a pacemaker-neuron-dependent manner which may contribute to specific functions such as different sensitivities to entraining cues.

Isolation and Analysis of the Yeast Mutant Gene, soo1-1, which Confers the Defect in $\beta$-1,3-glucan Biosynthesis (효모의 베타-1,3-글루칸 생합성 결함을 초래하는 돌연변이 유전자(soo1-1)의 분리 및 분석)

  • 이재준;이동원;김기현;박희문
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.42-48
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    • 2001
  • Allele rescue and sequence analysis of soo1-1 allele in Saccharomyces cerevisiae mutant LP0353 revealed that soo1-1 is identical to the previously reported ret1-1 allele, which has a base substitution of A for $G^{681}$ leading to an amino acid substitution of aspartic acid for $glycine^{227}$ in Soolp. However, it was revealed that the addition of osmotic stabilizer, such as 1.2M sorbitol can rescue the temperature sensitive phenotype of the ret1-1 mutant and that the soo1-1/ret1-1 mutation may confer defects in post-translational modification of proteins involved in the yeast cell wall biogenesis. Evidence for a putative role of 5th WD40 domain of the Soo1p/$\alpha$-COP in the construction and maintenance of cell walls was also presented by complementation test with deletion constructs of the SOOl.

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Isolation of Temperature-sensitive Mutant Escherichia coli YrdC Involved in Universal t6 A tRNA Synthesis (대장균에서 t6 A tRNA의 생합성에 관여하는 필수 단백질 YrdC의 온도 민감형 돌연변이 분리)

  • Hwang, Jihwan
    • Journal of Life Science
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    • v.28 no.2
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    • pp.257-264
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    • 2018
  • The YrdC superfamily is a group of proteins that are highly conserved in almost all organisms sequenced so far. YrdC in Escherichia coli was suggested to be involved in ribosome biogenesis, translation termination, cold adaptation, and threonylcarbamoyl adenosine formation in tRNA. In this study, to unambiguously demonstrate that yrdC is essential in E. coli, we constructed two yrdC mutant strains of E. coli and examined their phenotypes. In the temperature-sensitive yrdC mutant strain, cell growth stopped almost immediately under nonpermissive conditions and it appeared to accumulate 16S ribosomal RNA precursors without significant accumulation of 30S ribosomal subunits. We also cloned yeast and human homologs and demonstrated that they complement the E. coli yrdC-deletion strain. By mutational study, we demonstrated that the concave surface in the middle of the YrdC protein plays an important role in E. coli, yeast, and human versions. By comparison of two yrdC-deletion strains, we also unambiguously demonstrated that yrdC is essential for viability in E. coli and that the functions of its yeast and human homologs overlap with that of E. coli YrdC.