• 제목/요약/키워드: wild type

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참김 (Porphyra tenera Kjellman) 색소 및 형태변이체의 순계주 분리 및 특성 (Isolation and Characterization of Pure lines of Pigmentation and Morphological Mutants in Porphyra tenera Kjellman (Bangiales, Rhodophyta))

  • 황미숙;김승오;이영순;박은정;김성철;하동수;공용근;백재민;최한구
    • 한국수산과학회지
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    • 제43권5호
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    • pp.495-502
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    • 2010
  • Pure lines were isolated from young gametophytic blades of pigmentation and morphological mutants in Porphyra tenera. Growth, blade-shape and photosynthetic pigment content of pure lines were compared with the wild type. Growth of blade length in the wild type (W, R-B), with round shape and brown color, was fastest at $5{\sim}10^{\circ}C$ and became slower as temperature increased. The blade-shape of the wild type changed from linear to round as temperature increased. The green type (R-G), with round shape and green color, showed slower growth, and the red type (R-R) 'with round shape and red color' showed faster growth than the wild type. The blade-shapes of the green and red types changed from elliptical or linear to round as temperature increased. The phycoerythrin (PE) / phycocyanin (PC) ratio of the green type was markedly lower and the PE/PC ratio of the red type was markedly higher than that of the wild type. The linear type (L-B), with liner shape and brown color, showed faster growth in blade length than the wild type at $10{\sim}20^{\circ}C$ and maintained its linear shape at $5{\sim}15^{\circ}C$. The content of photosynthetic pigments of the linear type was similar to that of the wild type. Each of the pure lines of pigmentation and morphological mutants that were isolated in the present study showed particular patterns in growth, blade-shape and photosynthetic pigment composition. Therefore, they are expected to be useful as new varieties by themselves and to be available for breeding and biotechnological studies.

Xanthomonas campestris pv. campestris의 병원성 관련 형질 탐색에 관한 연구 (Molecular Approaches to Evaluate the Role of Some Genes Required for Plant Pathogenicity of Xanthomonas campestris pv. campestris)

  • 배동원;윤한대;김희규
    • 한국식물병리학회지
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    • 제13권3호
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    • pp.172-178
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    • 1997
  • 십자화과 작물에 발생하는 검은썩음병(Black rot or Black vein of crucifer)의 병원성 세균인 Xanthomonas campestris pv. crucifer)의 병원성 세균인 Xanthomonas campestris pv. campestris를 분리, 동정하고 병원성을 검정하였다. 이 X. c. pv. campestris 는 3가지 종의 Chinese cabbage에 병원성을 나타내었고, 병원성과 관련된 특성을 결정하기 위하여 Tn5 mutagenesis를 실시 cellulase negative mutant를 선발하여 병원성 검정하였다. 선발된 cellulase negative mutant를 배추에 분무 접종하여 광학 현미경과 전자현미경으로 관찰한 결과 cellulase negative mutant는 wild type와 함께 기공표면과 기공하부조직에서 정착하였지만 그 밀도는 낮았다. 반면 접종 24시간 이후 wild type은 기공표면과 기공하부조직이 lysis되기 시작하여 48시간 이후에는 병원성의 진전으로 보다 많이 lysis되었다. 6일 후, wild type은 cellulase활성에 의해 식물체 조직에서 높은 증식력을 보이며 조직을 lysis 시키고 또한 조직 깊숙이 침입, 정착하는 것을 관찰하였다. 이 결과로 X. c. pv.c campestris의 cellulase는 병원성에 관여하는 중요한 요인으로 생각된다.

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Crystallization and X-Ray Crystallographic Studies of Wild-Type and Mutant Tryptophan Synthase α-Subunits from Escherichia coli

  • Jeong, Mi Suk;Jang, Se Bok
    • Molecules and Cells
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    • 제19권2호
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    • pp.219-222
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    • 2005
  • The a-subunit of Escherichia coli tryptophan synthase (${\alpha}TS$), a component of the tryptophan synthase ${\alpha}_2{\beta}_2$ complex, is a monomeric 268-residues protein (Mr = 28,600). ${\alpha}TS$ by itself catalyzes the cleavage of indole-3-glycerol phosphate to glyceraldehyde-3-phosphate and indole, which is converted to tryptophan in tryptophan biosynthesis. Wild-type and P28L/Y173F double mutant ${\alpha}$-subunits were overexpressed in E. coli and crystallized at 298 K by the hanging-drop vapor-diffusion method. X-ray diffraction data were collected to $2.5{\AA}$ resolution from the wild-type crystals and to $1.8{\AA}$ from the crystals of the double mutant, since the latter produced better quality diffraction data. The wild-type crystals belonged to the monoclinic space group C2 ($a=155.64{\AA}$, $b=44.54{\AA}$, $c=71.53{\AA}$ and ${\beta}=96.39^{\circ}$) and the P28L/Y173F crystals to the monoclinic space group $P2_1$ ($a=71.09{\AA}$, b=52.70, $c=71.52{\AA}$ and ${\beta}=91.49^{\circ}$). The asymmetric unit of both structures contained two molecules of ${\alpha}TS$. Crystal volume per protein mass ($V_m$) and solvent content were $2.15{\AA}^3\;Da^{-1}$ and 42.95% for the wild-type and $2.34{\AA}^3\;Da^{-1}$ and 47.52% for the double mutant.

Role of Surface Protective Antigen A in the Pathogenesis of Erysipelothrix rhusiopathiae Strain C43065

  • Borrathybay, Entomack;Gong, Feng-juan;Zhang, Lei;Nazierbieke, Wulumuhan
    • Journal of Microbiology and Biotechnology
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    • 제25권2호
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    • pp.206-216
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    • 2015
  • To clarify the role of surface protective antigen A (SpaA) in the pathogenesis of Erysipelothrix rhusiopathiae C43065 (serotype 2), the spaA deletion mutant of E. rhusiopathiae ${\Delta}spaA$ was constructed by homologous recombination. The virulence of the ${\Delta}spaA$ mutant decreased more than 76-fold compared with that of the wild-type strain C43065 in mice. The mutant strain was sensitive to the bactericidal action of swine serum, whereas the wild-type strain was resistant. The adhesion of wild-type strain to MEF cells was inhibited significantly by treatment with rabbit antiserum against recombinant SpaA (rSpaA) as compared with the treatment with normal rabbit serum, but the mutant strain was not affected. The mutant strain was readily taken up by mouse peritoneal macrophages in the normal rabbit serum, whereas the wild-type strain was resistant. Whereas the rabbit antiserum against rSpaA promoted the phagocytosis of wild-type strain by macrophages, the mutant strain was not affected. In addition, mice vaccinated with the formalin-killed mutant strain were provided 40% protection against challenge by the homologous virulent strain as compared with those with wild-type strain, NaOH-extracted antigen, or rSpaA, which provided more than 80% protection against the same infection. These suggested that SpaA has an important role in the pathogenesis of E. rhusiopathiae infection and could be a target for vaccination against swine erysipelas.

Spectral Studies of Conformational Change at the Active Site of Mutant O-acetylserine Sulfhydrylase-A (C43S)

  • Park, Joon-Bum;Kim, Sung-Kun;Yoon, Moon-Young
    • BMB Reports
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    • 제29권1호
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    • pp.32-37
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    • 1996
  • The cysteine 43, potentially important in the activity of O-acetylserine sulfhydrylase (OASS) from Salmonella typhimurium, has been changed to serine. This mutant enzyme (C43S) has been studied in order to gain insight into the structural basis for the binding of inhibitor, substrate and product. UV-visible spectra of C43S exhibit the same spectral change in the presence of OAS as that observed with wild type enzyme, indicating C43S will form an ${\alpha}$-aminoacrylate Schiff base intermediate. At pH 6.5, however, the deacetylase activity of C43S is much higher than wild type enzyme indicating that cysteine 43 plays a role in stabilizing the ${\alpha}$-aminoacrylate intermediate. The fluoroscence spectrum of C43S exhibits a ratio of emission at 340 to 502 nm of 16.9, reflecting the lower fluorescence of PLP and indicating that the orientation of cofactor and tryptophan are different from that of the wild type enzyme. The emission spectrum of C43S in the presence of OAS gives two maxima at 340 and 535 nm. The 535 nm emission is attributed to the fluoroscence of the ${\alpha}$-aminoacrylate intermediate. The visible circular dichroic spectrum was similar to wild type enzyme, but the negative effect observed at 530~550 nm and the molar ellipicity values for the mutant are decreased by about 50% compared to wild type enzyme. The circular dichroic and fluoroscence studies suggest binding of the cofactor is less asymmetric in C43S than in the wild type enzyme.

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Contribution of the murI Gene Encoding Glutamate Racemase in the Motility and Virulence of Ralstonia solanacearum

  • Choi, Kihyuck;Son, Geun Ju;Ahmad, Shabir;Lee, Seung Yeup;Lee, Hyoung Ju;Lee, Seon-Woo
    • The Plant Pathology Journal
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    • 제36권4호
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    • pp.355-363
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    • 2020
  • Bacterial traits for virulence of Ralstonia solanacearum causing lethal wilt in plants were extensively studied but are not yet fully understood. Other than the known virulence factors of Ralstonia solanacearum, this study aimed to identify the novel gene(s) contributing to bacterial virulence of R. solanacearum. Among the transposon-inserted mutants that were previously generated, we selected mutant SL341F12 strain produced exopolysaccharide equivalent to wild type strain but showed reduced virulence compared to wild type. In this mutant, a transposon was found to disrupt the murI gene encoding glutamate racemase which converts L-glutamate to D-glutamate. SL341F12 lost its motility, and its virulence in the tomato plant was markedly diminished compared to that of the wild type. The altered phenotypes of SL341F12 were restored by introducing a full-length murI gene. The expression of genes required for flagella assembly was significantly reduced in SL341F12 compared to that of the wild type or complemented strain, indicating that the loss of bacterial motility in the mutant was due to reduced flagella assembly. A dramatic reduction of the mutant population compared to its wild type was apparent in planta (i.e., root) than its wild type but not in soil and rhizosphere. This may contribute to the impaired virulence in the mutant strain. Accordingly, we concluded that murI in R. solanacearum may be involved in controlling flagella assembly and consequently, the mutation affects bacterial motility and virulence.

Symbiotic Effectiveness of Bradyrhizobium japonicum USDA 110 in Supernodulating Soybean Mutant SS2-2

  • Lestari Puji;Van Kyujung;Kim Moon Young;Lee Suk-Ha
    • 한국작물학회지
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    • 제50권2호
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    • pp.125-130
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    • 2005
  • In the absence of exogeneous nitrogen supply, evaluation of a symbiosis effectiveness of Bradyrhizobium japonicum USDA 110 in a supernodulating soybean mutant, SS2-2, its wild type, Sinpaldalkong 2, and control genotype, Jangyeobkong, was conducted in this study. Nodules in SS2-2 were initially white and similar to its wild type, Sinpaldalkong 2. At the late stage, the wild type nodules became dark pinkish by maturation, by contrast, mature nodules in SS2-2 remained light green to pinkish, indicating a lack of leghemoglobin. Tap root length was short in nodulated symbiotic SS2-2 than that of its wild type and the control genotype. Nodulated root length and nodule density on root length were significantly increased by B. japonicum inoculation, but no significant increase was observed on root length and percentage of nodulation to total root length. Regardless of Bradyrhizobium inoculation, SS2-2 showed higher nodule dry weight and higher acetylene reduction activity (ARA) when compared with its wild type and the control genotype. Inoculation of B. japonicum leaded the increase of ARA in 47 days after planting (DAP), in part because of nodule development. Supernodulating mutant, SS2-2, less responded to B. japonicum induction in terms of nitrogen fixation and nodulation characteristics than its wild type. Thus, interaction of supernodulating soybean mutant with Bradyrhizobium had less symbiotically associated response than normal nodulating soybean.

Recognition of DNA by IHF : Sequence Specifficity Mediated by Residues That Do Not Contact DNA

  • Read, Erik K.;Cho, Eun Hee;Gardner, Jeffrey F.
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2001년도 Proceedings of 2001 International Symposium
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    • pp.35-39
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    • 2001
  • The Integration Host factor (IHF) of Escherichia coli is a small, basic protein that is required for a variety of functions including site-specific recombination, transposition, gene regulation, plasmid replication, and DNA packaging. It ,is composed of two subunits that are encoded by the ihfA ($\alpha$-subunit) and ihjB ($\beta$-subunit) genes. IHF binding sites are composed of three elements called the WATCAR, TTG, and poly (dAT) elements. We have characterized IHF binding to the H site of bacteriophage λ. We have isolated suppressors that bind to altered H' sites using a challenge phage selection. Two different suppressors were isolated that changed the adjacent $\alpha$P64 and $\alpha$K65 residues. The suppressors recognized both the wild-type site and a site with a change in the WATCAR element. Three suppressors were isolated at $\beta$-E44. These suppressors bound the wild-type and a mutant site with a T:A to A:T change (H44A) in the middle of the TIR element. Site-directed mutagenesis was used to make several additional changes at $\beta$E44. The wild-type and $\beta$E44D mutant could not bind the wild-type site but were able to bind the H44A mutant site. Other mutants with neutral, polar, or a positive charge at $\beta$E44 were able to repress both the wild-type and H44A sites. Examination of the IHF crystal structure suggests that the ability of the wild-type and $\beta$E44D proteins to discriminate between the T:A and A:T basepairs is due to indirect interactions. The $\beta$-E44 residue does not contact the DNA directly. It imposes binding specificity indirectly by interactions with residues that contact the DNA. Details of the proposed interactions are discussed.

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Ecophysiological Changes in a Cold Tolerant Transgenic Tobacco Plant Containing a Zinc Finger Protein (PIF1) Gene

  • Yun, Sung-Chul;Kwon, Hawk-Bin
    • 한국환경농학회지
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    • 제27권4호
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    • pp.389-394
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    • 2008
  • The ecophysiological changes occurring upon cold stress were studied using cold tolerant transgenic and wild-type tobacco plants. In a previous study, cold tolerance in tobacco was induced by the introduction of a gene encoding the zinc finger transcription factor, PIF1. Gas-exchange measurements including net photosynthesis and stomatal conductance were performed prior to, in the middle of, and after a cold-stress treatment of $1{\pm}2^{\circ}C$ for 96 h in each of the four seasons. In both transgenic and wild-type plants, gas-exchange parameters were severely decreased in the middle of the cold treatment, but had recovered after 2-3 h of adaptation in a greenhouse. Most t-test comparisons on gas-exchange measurements between the two plant types did not show statistical significance. Wild-type plants had slightly more water-soaked damage on the leaves than the transgenic plants. A light-response curve did not show any differences between the two plant types. However, the curve for assimilation-internal $CO_2$ in wild-type plants showed a much higher slope than that of the PIF1 transgenic plants. This means that the wild-type plant is more capable of regenerating Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) and has greater electron transport capacity. In conclusion, cold-resistant transgenic tobacco plants demonstrated a better recovery of net photosynthesis and stomatal conductance after cold-stress treatment compared to wild-type plants, but the ecophysiological recoveries of the transgenic plants were not statistically significant.