• Title/Summary/Keyword: point mutants

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The Rice FON1 Gene Controls Vegetative and Reproductive Development by Regulating Shoot Apical Meristem Size

  • Moon, Sunok;Jung, Ki-Hong;Lee, Do-Eun;Lee, Dong-Yeon;Lee, Jinwon;An, Kyungsook;Kang, Hong-Gyu;An, Gynheung
    • Molecules and Cells
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    • v.21 no.1
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    • pp.147-152
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    • 2006
  • Most plant organs develop from meristems. Rice FON1, which is an ortholog of Clv1, regulates stem cell proliferation and organ initiation. The point mutations, fon1-1 and fon1-2, disrupt meristem balance, resulting in alteration of floral organ numbers and the architecture of primary rachis branches. In this study, we identified two knockout alleles, fon1-3 and fon1-4, generated by T-DNA and Tos17 insertion, respectively. Unlike the previously isolated point mutants, the null mutants have alterations not only of the reproductive organs but also of vegetative tissues, producing fewer tillers and secondary rachis branches. The mutant plants are semi-dwarfs due to delayed leaf emergence, and leaf senescence is delayed. SEM analysis showed that the shoot apical meristems of fon1-3 mutants are enlarged. These results indicate that FON1 controls vegetative as well as reproductive development by regulating meristem size.

Determination of subcellular localization of Betanodavirus B2

  • Kim, Yeong-Mi;Cha, Seung-Ju;Mun, Chang-Hun;Do, Jeong-Wan;Park, Jeong-U
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2006.05a
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    • pp.476-478
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    • 2006
  • To analyze subcellular localization of betanodavirus protein B2, a plasmid expressing Betanodavirus protein B2 fused to enhanced green fluorescent protein (EGFP-Nl) was constructed. The transient expression of full-length B2 fused to EGFP in GF cells confirmed the equal distribution of protein B2 between cytoplasm and nucleus. However, transfection of N-terminal half of the B2 revealed that this truncated form predominantly localized to the cytoplasm. By using several deletion mutants and point mutants, we determined the regions and/or motif responsible for the subcellular localization of betanodavirus.

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Resistance Mechanism of Enterococcus faecalis to LCB01-0371, a New Oxazolidinone (새로운 옥사졸리디논계 항균제 LCB01-0371에 대한 Enterococcus faecalis의 내성 기전)

  • Lee, Hyun-Hee;Lee, Su-Ro;Kwak, Jin-Hwan
    • YAKHAK HOEJI
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    • v.58 no.1
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    • pp.7-11
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    • 2014
  • To study the resistance mechanism of E. faecalis to LCB01-0371, several resistant mutants to LCB01-0371 or linezolid were isolated by step-wise selection. The frequency of spontaneous mutations resistant to LCB01-0371 was lower than that of linezolid in E. faecalis. The genetic variations in resistant mutants were analyzed by DNA sequencing of domain V of 23S rRNA in each mutant. The first-step mutant to LCB01-0371 had a G2576T point mutation in V domain of 23S rRNA. However, no resistant mutant to LCB01-0371 was isolated in second-step mutant selection.

Isolation of Mycoplasma pneumoniae and Antimicrobial Susceptibilities of the Isolates(III) (Mycoplasma pneumoniae의 분리 및 항생제 감수성 검사(III))

  • Chang Myung-Woong;Kim Kwang-Hyuk;Park In-Dal;Song Gap-Young;Kim Sung-Won;Lee Eun-young;Kim Moon-Chan;Cho Myung-Hoon;Kim Kyu-Earn;Choi Choong-Eon;Park Seon Yeong;Jo Hyeon Jang
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.479-485
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    • 2005
  • The 994 throat swabs obtained from 688 adults and 306 children patients with respiratory diseases were examined for Mycoplasma pneumoniae infection by culture method. Antimicrobial susceptibilities of the resulting 123 M. pneumoniae isolates were evaluated by testing minimum inhibitory concentrations (MICs) of erythromycin, minocycline, tetracycline, josamycin, sparfloxacin, ofloxacin, and ciprofloxacin by a broth micro-dilution method. The erythromycin resistant strains of M. pneumoniae was determined above $1.0{\mu}g/ml$ of MIC for erythromycin. The erythromycin resistant strains of M. pneumoniae was confirmed resistant gene mutation of the portions of genes 23S rRNA (domain II and V), and ribosomal protein 14 and L22 by PCR amplified and their nucleotide sequenses were compared to those of the susceptible strain M129. The isolation rate of M. pneumoniae was $12.9\%$ (89/688) for the adults and $11.1\%$ (34/306) for the children. The $MICs_{90}$ of the M. pneumoniae isolates were $0.12{\mu}g/ml$ for minocycline, $0.25{\mu}g/ml$ for sparfloxacin, $0.5{\mu}g/ml$ for ciprofloxacin, ofloxacin, and tetracycline, respectively, and $2.0{\mu}g/ml$ for josamycin and erythromycin, respectively. The isolation rate of erythromycin resistant M. pneumoniae from patients was $49.4\%\;(44/89)$ for the adults, $47.1\%\;(16/34)$ for children, and $48.8\%\;(60/123)$ for the total. No mutation could be detected in the ribosomal protein L22 region, but all strains were mutated in the ribosomal protein L4 as two point mutation M144V. Two point mutations in domain V of 23S rRNA were selected in the presense of erythromycin resistant M. pneumoniae isolates, such as one strain was G2057C mutant, two strains were A2059C mutants, three strains were C2611G mutants, four strains were A2058C mutants, five strains were A2058T mutants, twenty strains were A2059G mutants, and twenty-five strains were A2058G mutants, respectively. These results show that erythromycin was not the most active compound against M. pneumoniae infection in Korea and clinical studies of macrolides in human patients are demanded.

Cloning and Site-Directed Mutagenesis of Musca domestica Acetylcholinesterase for Enhancing Sensitivity to Organophosphorus and Carbamate Insecticides

  • Kim, Chung-Sei;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1760-1772
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    • 2006
  • Mature acetylcholinesterase (AChE) gene (gm, 1,836 bp) was cloned from the housefly and successfully expressed in the E. coli CodonPlus (DE3) RIL system (GM-E, 72 kDa) with a yield of 1,630 mU/g fresh cells. Using the gm, 10 kinds of mutants were constructed and expressed for enhancing sensitivity to insecticides. The sensitivity of these mutants to five kinds of organophosphate (OP) and three carbamate insecticides was investigated by measuring the apparent bimolecular inhibition constant ($k_i=k_2/K_d$). Surprisingly, the sensitivity of quadruple mutant IGFT was enhanced as much as 7-fold for acephate, 164-fold for demeton-S-methyl, 484-fold for dichlorvos, 523-fold for edifenphos, 30-fold for ethoprophos, 30-fold for benfuracarb, 404-fold for carbaryl, and 107-fold for furathiocarb, compared with that of GM-E, although the sensitivity of each single point mutant was slightly increased. These mutational studies indicated that (i) contradictory to Walsh et al. [39], the residue 327 is the important key residue for enhancing sensitivity as much as the residue 262, (ii) the residue 82 and additional residues of 234, 236, and 585 are also important, and (iii) sensitivity was cooperatively accelerated as the number of strategic mutations increased.

In vitro Mouse Lymphoma Thymidine Kinase (tk+/-) Gene Forward Mutation Assay in Mammalian cells (포유동물세포의 Forward Mutation을 지표로 한 Mouse Lymphoma Thymidine Kinase (tk+/-) Gene Assay)

  • 류재천;김경란;최윤정
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.7-13
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    • 1999
  • The mouse lymphoma thymidine kinase (tk+/-) gene assay (MOLY) using L5178Y tk+/- mouse lymphoma cell line is one of the mammalian forward mutation assays. It is well known that MOLY has many advantages and more sensitive than the other mammalian forward mutation assays such as x-linked hyposanthine phosphoribosyltransferase (hprt) gene assay. The target gene of MOLY is a heterozygous tk+/- gene located in 11 chromosome of L5178Y tk+/- cell, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. MOLY has relatively short expression time (2-3 days) compared to 1 week of hprt gene assay. MOLY can also induce relatively high mutant frequency so a large number of events can be recorded. The bimodal distribution of colony size which may indicate gene mutation and chromosome breakage potential of chemicals according to mutation scale such as large normal-growing mutants and small slow-growing mutants can be observed in this assay. The statistical analysis of data can be performed using the MUTANT program developed by York Electronic Research in association with Hazelton as recommended by the UKEMS (United Kingdom Environmental Mutagen Society) guidelines. This report reviewed MOLY using the microtiter cloning technique (microwell assay).

Induction and Chatacterization of pKM101 Mutants in Salmonella typhimurium (Salmonella typhimurium내로의 pKM101 돌연변이체의 유도와 그 특성에 관한 연구)

  • 백형석;강수형;이세영
    • Korean Journal of Microbiology
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    • v.20 no.2
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    • pp.89-97
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    • 1982
  • Mutants of plasmid pKM101 modified to enhance mutagenesis were induced and characterized in Salmonella typhimurium. The pKM101 mutant plasmid were transferred normally and stably maintained in cells. They had modified in their ability (i) to enhance the reversion of both point and frameshift mutations, (ii) to protect the cell against UV-irradiation and chemical mutagen treatment, (iii) of ampicillin resistance. A similar modification in enhancement of reversion was also observed in a $uvrB^-$ strains. These results indicated that mutator effect of pKM101 was coded by one plasmid gene.

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Cell Cycle Regulation in the Budding Yeast

  • Nguyen, Cuong;Yoon, Chang-No;Han, Seung-Kee
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.278-283
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    • 2005
  • Cell cycle is regulated cooperatively by several genes. The dynamic regulatory mechanism of protein interaction network of cell cycle will be presented taking the budding yeast as a sample system. Based on the mathematical model developed by Chen et at. (MBC, 11,369), at first, the dynamic role of the feedback loops is investigated. Secondly, using a bifurcation diagram, dynamic analysis of the cell cycle regulation is illustrated. The bifurcation diagram is a kind of ‘dynamic road map’ with stable and unstable solutions. On the map, a stable solution denotes a ‘road’ attracting the state and an unstable solution ‘a repelling road’ The ‘START’ transition, the initiation of the cell cycle, occurs at the point where the dynamic road changes from a fixed point to an oscillatory solution. The 'FINISH' transition, the completion of a cell cycle, is returning back to the initial state. The bifurcation analysis for the mutants could be used uncovering the role of proteins in the cell cycle regulation network.

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Phenotypes of Integrase-Mutated Human Immunodeficiency Virus Type-1(HIV-1)

  • ;Chris M. Farnet;William A. Haseltine
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.92-92
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    • 1993
  • Point mutations in a highly conserved central region of the HIV-1 integrase protein were analyzed for their effects on viral replication and virion morphogenesis. Conservative amino acid replacements of two amino acid residues invariant un retroviral integrases, D116 and E152 of HIV-1, as well as the highly conserved amino acid S147, completely blocked viral replication in two CD4$\^$+/ human T cell lines. Mutation of four other highly conserved amino acids in the region had no detectable effect on viral replication, while Mutations at two positions, N117 and Y143, resulted in viruses with a delayed replication phenotype. Characteristic and reproducible defects id virion core structure were observed by electron microscopic analysis of sore of the replication defective integrase point mutants, indicating that mutant integrase proteins can interfere with the process of virion core maturation.

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Studies on Mild Mutants of Tabacco Mosaic Virus I. Induction of Mild Mutants and Their Characteristics (약독 담배 모자이크 바이러스 I. 약독바이러스의 유기 및 특성)

  • Choi Jang Kyung;Son Kyung Ok
    • Korean Journal Plant Pathology
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    • v.1 no.1
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    • pp.3-11
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    • 1985
  • Three mild mutant strains of tobacco mosaic virus (TMV) were isolated from Nicotiana tabacum var. Samsun incubated at $38^{\circ}C$ for 10 days after inoculation with a wild type of TMV-OM strain. They were designated into Tg 5272, Tw 227 and Tw 333. All mild strains could be distinguished from TMV-OM by their reactions on different indicator plants. The mild strains induced the mild mottling without distinct symptoms, whereas the wild strain produced severe mosaic, rugose and stunting on tobacco and red pepper plants. Tw 227 and Tw 333 produced smaller necrotic spots than those of Tg 5272 and TMV-OM on N. glutinosa and Datura stramonium. The former two strains also produced ring spots and mosaic on Gomphrena globosa compared with necrotic spots by the latter strains. Three mild strains were serologically identical to TMV-OM. Their physical properties were thermal inactivation point $80-85^{\circ}C$, dilution end point between $10^{-4}\;and\;10^{-6}$, and longevity in vitro 7days or longer. Ultraviolet absorption spectra of purified preparations of the mild strains and TMV-OM were identical, with a minimum at 247nm, a maximum at 260nm, and a slight shoulder at 290nm. Electrophoresis of the strains in polyacrylamide-agarose gel showed that all the strains formed one major band and two minor bands, except for one minor band of Tw 333. However, when sodium dodecyl sulfate was added to the purified viruses before electrophoresis, each strain formed only one major band.

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