• 제목/요약/키워드: Wild type and mutants

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Selection of Nitrate-nonutilizing Mutants of Hypoxylon atropunctatum, A Fungal Pathogen on Oak Species

  • Chun, Se-Chul;Fenn, P.
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.247-251
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    • 2000
  • Latent infections of healthy-appearing oaks of Hypoxylon atropunctatum complicates field studies by interfering with inoculation experiments to follow pathogenesis, fungal development and reproduction of this canker rot fungus. Mutants with unique and easily scorable phenotypes would be useful for inoculation studies. There is a broad range in the capacity of wild-type isolates to utilize nitrate as a sole nitrogen sources. Several types of nitrate-nonutilization mutants (nit1, Nit3, NitM) were selected from nitrate-utilizing wild-type isolates. Also, a few mutants of Hypoxylon atropunctatum were selected that could only grow poorly on basal medium supplemented with various nitrogen sources and even on yeast extract agar. These unknown mutants need to be characterized further. Nit mutants of Hypoxylon atropunctatum were readily selected, grew well and were recovered after inoculation into oak stems. These results suggest that nit mutants could be useful for inoculation studies in trees that contain latent infections.

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Isolation and Characterization of Pre-$tRNA^{Val}$ Splicing Mutants of Schizosaccharomyces pombe

  • Hwang, Ku-Chan;Kim, Dae-Myung
    • Journal of Microbiology
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    • v.35 no.4
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    • pp.334-340
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    • 1997
  • A collection of 132 temperature sensitive (ts) mutants was generated by the chemical mutagenesis of Schizosaccharomyces pombe wild type strain and screened for tRNA splicing defects on Northern blots by hybridization with an oligonucleotide that recognizes the exon of the S. pombe tRNA^Val as a probe. We identidied 6 mutants which accumulate precursor $tRNA^{Val}$. Among them, 2 mutants exhibited remarkable morphological differences compared to wild type cells. One tRNA splicing mutant showed elongated cell shape in permissive as well as non-permissive cultures. The other mutant exhibited shortened cell morphology only in nonpermissive culture. The total RNA pattern in the splicing mutants appeared to be normal. Genetic analysis of four $tRNA^{Val}$ splicing mutants demonstrated that the mutation reside in different genes.

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Arabidopsis SIZ1 positively regulates alternative respiratory bypass pathways

  • Park, Bong-Soo;Kim, Sung-Il;Song, Jong-Tae;Seo, Hak-Soo
    • BMB Reports
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    • v.45 no.6
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    • pp.342-347
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    • 2012
  • Plant mitochondria possess alternative respiratory pathways mediated by the type II NAD(P)H dehydrogenases and alternative oxidases. Here, E3 SUMO ligase was shown to regulate alternative respiratory pathways and to participate in the maintenance of carbon and nitrogen balance in Arabidopsis. The transcript abundance of the type II NAD(P)H dehydrogenases NDA2 and NDB2 and alternative oxidases AOX1a and AOX1d genes was low in siz1-2 mutants compared to that in wild-type. The addition of nitrate or ammonium resulted in a decrease or an increase in the expression of the same gene families, respectively, in both wild-type and siz1-2 mutants. The amount of free sugar (glucose, fructose and sucrose) was lower in siz1-2 mutants than that in wild-type. These results indicate that low nitrate reductase activity due to the AtSIZ1 mutation is correlated with an overall decrease in alternative respiration and with a low carbohydrate content to maintain the carbon to nitrogen ratio in siz1-2 mutants.

Selective Isolation and Characterization of Schwanniomyces castellii Mutants with Increased Production of a-Amylase and Glucoamylase

  • Ryu, Yeon-Woo
    • Journal of Microbiology and Biotechnology
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    • v.3 no.2
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    • pp.95-98
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    • 1993
  • This study was carried out to isolate and characterize the mutant strains of Schwanniomyces castellii NRRL Y-2477. Mutants were prepared with the treatment of ethyl methane sulfonate. 2-deoxy-D-glucose resistant mutants were isolated and two mutants were selected based on their high production of amylolytic enzymes and their ability to ferment starch. The mutants selected had higher a-amylase and glucoamylase activities than the wild type strain from several other carbon sources. Especially, it was revealed that mutant strain M-9, when cultured in the presence of glucose as a sole carbon source, shows relatively high activities of a-amylase and glucoamylase compared to those of the wild type strain. In result, this mutant strain can be considered as a constitutive producer of amylolytic enzymes. To compare the ethanol production ability of wild type strain and of mutant strains selected, an alcohol fermentation was carried out using 100 g/l soluble starch. Mutant strain M-9 did not improve the direct alcohol fermentation of starch, despite its excellent amylolytic activities performance. On the other hand, mutant strain M-6 produced 37.9 g/l (4.8%, v/v) ethanol by utilizing about 82% of substrate.

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Isolation and Characterization of Benomyl-Resistant Mutants in an Entomopathogenic Fungus, Metarhizium anisopliae

  • Kim Soon Kee;Shim Hee Jin;Roh Jong Yul;Jin Byung Rae;Boo Kyung Saeng;Je Yeon Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.2
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    • pp.119-123
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    • 2005
  • Benomyl-resistant mutants of entomopathogenic fungus, Metarhizium anisopliae were isolated and their physiological characteristics were investigated. These militants were obtained spontaneously or by UV irradiation in benomyl-treated media. Four spontaneous (S-2, S-11, S-18, S-19) and four UV-induced (UV-4, UV-5, UV-19, UV-24) mutants, which grow stably and normally were selected. No significant differences in conidia or hyphal shape, conidia viability, mycelial biomass, or virulent to the diamondback moth were observed between the wild type and their mutants. But differently from the mycelial growth of other benomyl-resistant mutants which was slower than that of the wild type on a modified Czapek-Dox, SDAY, $4\%$ chitin, or $1\%$ skim milk medium, that in the spontaneous mutants, S-18 and S-19, did not show any difference from the wild type. Especially, S-18 and S-19 grew well at benomyl concentrations up to 50 times or higher than that which inhibits wild type proliferation. These results suggested that S-18 and S-19 could potentially be used with the fungicide, benomyl.

Isolation of Arthrospira platensis Mutants Producing High Lipid and Phycobiliproteins (지질과 phycobiliproteins 고생산성 Arthrospira platensis 변이주 분리)

  • Kim, Young-Hwa;Lee, Jae-Hwa
    • KSBB Journal
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    • v.27 no.3
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    • pp.172-176
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    • 2012
  • In this study, microalgae Arthrospira platensis (A. platensis) mutants induced by ethyl methane sulfonate (EMS) and further selection for resistance of cerulenin, a potent inhibitor of fatty acid synthase, were characterized. The mutants selected by $2{\mu}M$, $5{\mu}M$ and $10{\mu}M$ of cerulenin were designated EC2, EC5 and EC10, respectively. Under normal growth conditions, the mutants and parental strain exhibited similar growth pattern. The mutants of A. platensis showed enhanced lipid accumulation and phycobiliproteins (phycoerythrin, phycocyanin). The lipid content of mutants EC2 and EC5 was about 4.4 and 4.8-fold higher than wild type. The phycoerythrin and phycocyanin content of mutants EC2 and EC5 was increased about 1.5 and 6.9-fold and 1.4 and 3.8-fold, respectively, compared to the wild type. The chlorophyll and carotenoid content of mutants was slightly increased. The high lipid and pigment contents exhibited by A. platensis mutants would make an excellent candidate for the production of commercially interesting biologically active compounds.

Characteristics, Stability and Reisolation of nit Mutant of Fusarium oxysporum from Strawberry (딸기로부터 분리된 Fusarium oxysporum nit 변이주의 특성과 안정성 및 재분리)

  • 신동범;홍연규;조재민
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.583-588
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    • 1998
  • This study was conducted to investigate the potential of nitrate-nonutilizing mutants (nit mutants) in ecological studies of Fusarium disease of strawberry. Nit mutants of Fusarium oxysporum from strawberry were easily formed on chlorate-containing media. Nit mutants were assigned to three phenotypic classes, nit1, nit3, and NitM, on the basis of their growth on media containing one of the following five different nitrogen sources ; nitrate, nitrite, hypoxanthine, ammonium and uric acid. Frequency of nit mutation and proportion of three phenotypes of nit mutants depended on the isolate. Mutation rate was 45.6% and ranged from 15.0% to 95.0%. The frequency of nit1 mutants was higher than that of nit3 or NitM. The complementary reaction between nit1 and NitM was higher than that of other combination. There has been no complementary response observed between nit3 and nit3. The nit mutants showed similar growth pattern as the that of wild type isolate on potato sucrose agar and potato sucrose liquid media. Most of the mutants retained pathogenicity, and maintained their phenotypes even after two year preservation through subculture on slanted PSA at room temperature. Nit mutants were selctively isolated from infested soil and infected plants on the selective medium (MMCPA) containing potassium chlorate with their original phenotypes, while naturally occurring isolates of Fusarium oxysporum were not grow on the medium. On the contrary, nit mutants showed very slight growth on the medium (MMPA) containing nitrate as a sole nitrogen source, and therefore could be distinguished from wild type isolate.

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Functional Abnormalities of HERG Mutations in Long QT Syndrome 2 (LQT2)

  • Hiraoka, Masayasu
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.5
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    • pp.367-371
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    • 2001
  • The chromosome 7-linked long QT syndrome (LQT2) is caused by mutations in the human ether-a- go-go-related gene (HERG) that encodes the rapidly activating delayed rectifier $K^+$ current, $I_{Kr},$ in cardiac myocytes. Different types of mutations have been identified in various locations of HERG channel. One of the mechanisms for the loss of normal channel function is due to membrane trafficking of channel protein. The decreased channel function in some deletion mutants appears to be due to loss of coupling with wild type HERG to form the functional channel as the tetramer. Most of missense mutants with few exceptions could interact with wild type HERG to form functional tetramer and caused dominant negative suppression with co-injection with wild type HERG showing variable effects on current amplitude, voltage dependence, and kinetics of activation and inactivation. Two missense mutants at pore regions of HERG found in Japanese LQT2 (A614V and V630L) showed accentuated inward rectification due to a negative shift in steady-state inactivation and fast inactivation. One mutation in S4 region (R534C) produced a negative shift in current activation, indicating the S4 serving as the voltage sensor and accelerated deactivation. The C-terminus mutation, S818L, could not express the current by mutant alone and did not show dominant negative suppression with co-injection of equal amount of wild type cRNA. Co-injection of excess amount of mutant with wild type produced dominant negative suppression with a shift in voltage dependent activation. Therefore, multiple mechanisms are involved in different mutations and functional abnormality in LQT2. Further characterization with the interactions between various mutants in HERG and the regulatory subunits of the channels (MiRP1 and minK) is to be clarified.

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Isolation and Characterization of Regulatory Mutant for Cellulase Production from Trichoderma reesei QM 9414 (Trichoderma reesei QM 9414의 섬유소 분해 호소 생산을 위한 조절변이주의 분리 및 특성에 관한 연구)

  • Choi, Kun-Ho;Koo, Youn-Mo;So, Jae-Seong
    • The Korean Journal of Mycology
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    • v.26 no.1 s.84
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    • pp.127-133
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    • 1998
  • Two regulatory mutants of Trichoderma reesei QM 9414 were isolated by treatment with N-methyl-N'-nitro-N-nitrosoguanidine, and the effects of various inducers on the carboxymethyIcellulose (CMC) and filter paper (FP) production were investigated. Induction of CMCase and FPase production of mutants was shown higher level than wild type strain in 1% lactose minimal broth. When induced by glucose, wild type showed glucose-repression for CMCase and FPase production and mutants showed glucose-derepression. Mutant 1 showed 8.38 fold higher CMCase activity and 5.68 fold higher FPase activity than wild type stain. Mutant 2 showed about 8.42 fold higher CMCase activity and 5.41 fold higher FPase activity than wild type strain. Enzyme activities from the mutants and wild type had the same optimum pH of 4.8 and optimum temperature of $60^{\circ}C$.

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Cell-Surface Loss of Constitutive Activating and Inactivating Mutants of Eel Luteinizing Hormone Receptors

  • Byambaragchaa, Munkhzaya;Choi, Seung-Hee;Kim, Dong-Wan;Min, Kwan-Sik
    • Development and Reproduction
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    • v.25 no.4
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    • pp.225-234
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    • 2021
  • The present study aimed to investigate the mechanism of cell surface receptor loss by two constitutively activating mutants (designated L469R, and D590Y) and two inactivating mutants (D417N and Y558F) of the luteinizing hormone receptor (LHR) in the Japanese eel Anguilla japonica, known to naturally occur in human LHR transmembrane domains. We investigated cell surface receptor loss using an enzyme-linked immunosorbent assay in HEK 293 cells. The expression level of wild-type eel LHR was considered to be 100%, and the expression levels of L469R and D417N were 97% and 101%, respectively, whereas the expression levels of D590Y and Y558F slightly increased to approximately 110% and 106%, respectively. The constitutively activating mutants L469R and D590Y exhibited a decrease in cell surface loss in a manner similar to that of wild-type eel LHR. The rates of loss of cell surface agonist-receptor complexes were observed to be very rapid (2.6-6.2 min) in both the wild-type eel LHR and activating mutants. However, cell surface receptor loss in the cells expressing inactivating mutants D417N and Y558F was slightly observed in the cells expressing inactivating mutants D417N and Y558F, despite treatment with a high concentration of agonist. These results provide important information on LHR function in fish and the regulation of mutations of highly conserved amino acids in glycoprotein hormone receptors.