• Title/Summary/Keyword: double mutant

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Regulation of Ethylene Biosynthesis in Phytochrome Mutants of the Arabidopsis Root (Arabidopsis 피토크롬 돌연변이체에서 ethylene 생합성 조절 작용)

  • Park, Ji-Hye;Kim, Soon-Young
    • Journal of Life Science
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    • v.22 no.4
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    • pp.559-564
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    • 2012
  • In order to investigate the effect of phytochromes on the regulation of ethylene biosynthesis, we measured the ethylene production and the activities of enzymes involved in ethylene biosynthesis using phytochrome mutants such as $phyA$, $phyB$, and $phyAB$ of Arabidopsis. The ethylene production was decreased in mutants grown in white light. In particular, double mutants showed a 37% decrease compared to the wild type in ethylene production. When Arabidopsis roots were grown in the dark, mutants did not show a decrease in ethylene production; however, production was significantly decreased in the double mutant grown in red light. Only $phyB$ did not show the decrease in the ethylene production in far-red light. Unlike the ACO activities, the ACS activities of mutants showed the same pattern as the ethylene production under several light conditions. The results of ACS activities confirmed the expression of the ACS gene by RT-PCR analysis. The decrease of ethylene production in mutants was due to the lower activity of ACC synthase, which converts the S-adenosyl-L-methionine (AdoMet) to 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene. These results suggested that both phytochrome A and B play an important role in the regulation of ethylene biosynthesis in Arabidopsis roots in the conversion step of AdoMet to ACC, which is regulated by ACS.

Genetic Polymorphisms in Plasmodium vivax Dihydrofolate Reductase and Dihydropteroate Synthase in Isolates from the Philippines, Bangladesh, and Nepal

  • Thongdee, Pimwan;Kuesap, Jiraporn;Rungsihirunrat, Kanchana;Dumre, Shyam Prakash;Espino, Effie;Noedl, Harald;Na-Bangchang, Kesara
    • Parasites, Hosts and Diseases
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    • v.53 no.2
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    • pp.227-232
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    • 2015
  • Genetic polymorphisms of pvdhfr and pvdhps genes of Plasmodium vivax were investigated in 83 blood samples collected from patients in the Philippines, Bangladesh, and Nepal. The SNP-haplotypes of the pvdhfr gene at the amino acid positions 13, 33, 57, 58, 61, 117, and 173, and that of the pvdhps gene at the positions 383 and 553 were analyzed by nested PCR-RFLP. Results suggest diverse polymorphic patterns of pvdhfr alone as well as the combination patterns with pvdhps mutant alleles in P. vivax isolates collected from the 3 endemic countries in Asia. All samples carried mutant combination alleles of pvdhfr and pvdhps. The most prevalent combination alleles found in samples from the Philippines and Bangladesh were triple mutant pvdhfr combined with single mutant pvdhps allele and triple mutant pvdhfr combined with double wild-type pvdhps alleles, respectively. Those collected from Nepal were quadruple mutant pvdhfr combined with double wild-type pvdhps alleles. New alternative antifolate drugs which are effective against sulfadoxine-pyrimethamine (SP)-resistant P. vivax are required.

The screening & characterization of super-producing recombinant Hansenula polymorpha mutant

  • Gang, Hwan-Gu;Park, Hyeong-Su;Lee, Chung-Yeol;Yu, Byeong-Il;Yu, Eun-Jeong;Lee, Seon;Hwang, Seon-Deok;Gang, Hyeon-A;Lee, Sang-Gi
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.684-687
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    • 2000
  • The super-producing recombinant H.polymorpha mutant is obtained by double membrane screening technique combined with optimum mutation method. The characterization of mutant is carried out to find the change of mutant in m-RNA level, cell wall leakage, protease level and methanol utilization metabolic flux. The change of these properties of mutant was figured out.

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Characteristics of the Resistance Phenotypes by Inducible Resistance Gene ermK and Its Terminator Region Mutants (MLS계 항생제에 대한 유도내성 유전자 ermK 및 그 돌연변이체의 유도내성 표현형)

  • Choi, Sung-Sook;Choi, Eung-Chil
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.533-537
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    • 1997
  • The characteristics of the resistance phenotypes of Bacillus subtilis having ermk and its terminator region mutants were determined. Wild type ermK(pEC101) and pECMT109(methylase SD-region mutant) showed typical inducible resistance phenotype. pECMF1(terminator1 region mutant) and pECMT2(terminator2 region mutant) showed constitutive resistance to Kitasamycin but inducible resistance to tylosin. In contrast, pECMT3(terminator1 and terminator2 double mutant) and pECMT309(terminator1, terminator2 and methylase SD region triple mutant) showed constitutive resistance both to kitasamycin and tylosin.

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Improvement of Photoheterotrophic $H_2$ production of Rhodobacter sphaeroides by Removing Ammonium Ion Effect Exerted on Nitrogenase (Rhodobacter sphaeroides의 질소고정효소에 미치는 암모니움 이온 효과 제거를 통한 수소생성 증진)

  • Jin, Sang-Hoon;Kim, Mi-Sun;Lee, Jeong-Kug
    • KSBB Journal
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    • v.20 no.6
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    • pp.418-424
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    • 2005
  • Photoheterotrophic evolution of molecular hydrogen by Rhodobacter sphaeroides is mediated by nitrogenase that is regulated transcriptionally and post-translationally by ammonium ion. Two PII-like proteins, GlnB and GlnK, play key roles in mediating inhibition and repression of nitrogenase in the presence of ammonium ion. glnB and glnK of R. sphaeroides were interrupted to abolish the ammonium ion effect controlling nitrogenase. Ammonium ion effect was still observed in mutant having an interruption in either glnB or glnK. However, the nitrogenase activity of glnB-glnK double mutant is not affected by ammonium ion. $H_2$ evolution was improved by increasing gene dosages of nitrogenase-coding genes, nifHDK in trans in glnB-glnK double mutant.

Interaction of brassinosteroids and cytokinin in modulating light mediated signaling in Arabidopsis

  • Hwang, Indeok;Paudyal, Dilli P.;Cheong, Hyeonsook
    • Journal of Integrative Natural Science
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    • v.1 no.1
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    • pp.24-31
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    • 2008
  • Brassinosteroids (BRs) are a special class of plant steroid hormones that are essential for normal growth and development. Part of confusion is whether BRs are unique to plants, because they have overlapping physiological roles with other better-studied hormones and with physiological responses caused by light. In systems designed to assay for cytokinins, the effects of BRs vary. We measured hypocotyl length for testing the ability of brassinolide (BL) to rescue double mutant between det2 and the photoreceptor null mutant phytochrome B (phyB). PHYB involved in controlling hypocotyl elongation in increased concentration of BL whereas phyBdet2 double mutant just partially rescue to phyB in white and red light indicated the involvement of BRs in PHYB regulated cell elongation. BRs regulated hypocotyl growth was delayed by BAP, a cytokinin treatment but inhibitory effects of BAPs on hypocotyl growth was slightly recovered by BL. The result indicated that the mode of action of BR and cytokinin is independent or sequential in the downstream light-regulated response control on hypocotyl elongation and also light modulated the action of BR and cytokinin in some extent.

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Isolation of Oligosporogenous (Spo-Cry+)Mutant Containing Relatively tow Hemolytic Activity in $\delta$-Endotoxin from Bacillus thuringiensig subsp, israelensis (Bacillus thuringiensig subsp, israelensis의 용혈성 약화 및 무포자 변이주의 분리)

  • 배점순;김광현
    • Microbiology and Biotechnology Letters
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    • v.17 no.4
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    • pp.273-276
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    • 1989
  • The isolate SPO 3, an oligosporogenous and crystalliferous mutant, which was isolated from Bacillus thuringiensig subsp, israelensis by heat treatment at 42$^{\circ}C$ for 24 hit. The $\delta$-endotoxin of the mutant had a relatively low hemolytic activity on human red blood cells; the $\delta$-endotoxin of the mutant had 25 times less hemolytic activity compared to that of wild strain. The loss of 28 KDa hemolytic protein subunit in $\delta$-endotoxin of the mutant was confirmed by means of double immunodiffusiori, immunoelectrophoresis, and SDS-PAGE.

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Isolation and Characterization of Double Deletion Mutants of nsdD and veA or nsdC, the Genes Positive Regulating Sexual Development of Aspergillus nidulans (Aspergillus nidulans에서 유성분화 능동조절 유전자 nsdD와 veA 또는 nsdC의 이중결실돌연변이의 분리 및 분석)

  • Kim, Lee-Han;Kim, Ha-Eun;Han, Dong-Min
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.419-423
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    • 2013
  • Three genes, nsdD, nsdC, and veA are known to be necessary for positive regulating sexual development of Aspergillus nidulans. Since the mutants of those genes hardly form fruiting bodies in heterokaryons constructed by cross between two of them, it is difficult to isolate double mutants. In this work, double mutants of ${\Delta}nsdD$ ${\Delta}veA$ and ${\Delta}nsdD$ ${\Delta}nsdC$ were isolated using the characteristic of the nsdD deletion mutant that it could develop mature cleistothecia in hypoxic and low temperature culture condition. According to the phenotypes of double mutants, the nsdD gene controls the apical growth independently with veA or nsdC. Deletion of veA or nsdC was epistatic to nsdD deletion for pigment production. Conidia formation in submerged culture with lactose as sole carbon source was observed in ${\Delta}nsdD$ ${\Delta}nsdC$ double mutant implicating it to be unique phenotype of nsdC deletion.

Role of plastidic glucose transporter in source metabolism of Arabidopsis

  • Lee, Youn-Hyung;Hong, Soon-Won;Lee, Jang-Wook;Bhoo, Seong-Hee;Jeon, Jong-Seong;Hahn, Tae-Ryong
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.9-21
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    • 2005
  • To study the biochemical and physiological role of the plastidic glucose transporter (pGlcT) in carbohydrate metabolism, we characterized transgenic plants with mutations in the pGlcT gene (GT), gt-1 and gt-2, as well double mutants of GT and the maltose transporter (MEX1) and GT and the triose phosphate/phosphate translocator (TPT), GT and the cytosolic fructose-1,6-bisphosphatase gene (cFBP), and MEX1 and TPT, gt-1/mex2, gt-1/tpt-2, gt-1/cfbp-1, mex1-1/tpt-2, respectively. Compared to the wild type, all mutants except the gt-1/cfbp-1 mutant lines displayed higher starch accumulation and higher levels of maltose. Starch accumulation is due to a decrease in starch turnover, leading to an imbalance between the rates of synthesis and degradation. Sucrose levels of gt alleles were higher than those in wild-type plants during the light period, suggesting possible nightly supplementation via the maltose transport pathway to maintain proper carbohydrate partitioning in the plant leaves. The gt plants displayed less growth retardation than mex1-1 mutant and gt-1/mex2 double mutant displayed accumulativesevere growth retardation as compared to individual gt-1 and mex1-1 mutants, implying that the maltose transporter-mediated pathway is a major route for carbohydrate partitioning at night. The gt-1/tpt-2, mex1-1/tpt-2 and gt-1/cfbp-1 double mutants had retarded growth and low chlorophyll content to differing degrees, indicating that photosynthetic capacity had diminished. Interestingly, the gt-1/tpt-2 line displayed a glucose-insensitive phenotype and higher germination rates than wild type, suggesting its involvement not only in carbon partitioning, but also in the sugar signaling network of the pGlcT and TPT.

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Asn-Linked Glycosylation Contributes to Surface Expression and Voltage-Dependent Gating of Cav1.2 Ca2+ Channel

  • Park, Hyun-Jee;Min, Se-Hong;Won, Yu-Jin;Lee, Jung-Ha
    • Journal of Microbiology and Biotechnology
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    • v.25 no.8
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    • pp.1371-1379
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    • 2015
  • The Cav1.2 Ca2+ channel is essential for cardiac and smooth muscle contractility and many physiological functions. We mutated single, double, and quadruple sites of the four potential Asn (N)-glycosylation sites in the rabbit Cav1.2 into Gln (Q) to explore the effects of Nglycosylation. When a single mutant (N124Q, N299Q, N1359Q, or N1410Q) or Cav1.2/WT was expressed in Xenopus oocytes, the biophysical properties of single mutants were not significantly different from Cav1.2/WT. In comparison, the double mutant N124,299Q showed a positive shift in voltage-dependent gating. Furthermore, the quadruple mutant (QM; N124,299,1359,1410Q) showed a positive shift in voltage-dependent gating as well as a reduction of current. We tagged EGFP to the QM, double mutants, and Cav1.2/WT to chase the mechanisms underlying the reduced currents of QM. The surface fluorescence intensity of QM was weaker than that of Cav1.2/WT, suggesting that the reduced current of QM arises from its lower surface expression than Cav1.2/WT. Tunicamycin treatment of oocytes expressing Cav1.2/WT mimicked the effects of the quadruple mutations. These findings suggest that Nglycosylation contributes to the surface expression and voltage-dependent gating of Cav1.2.