• Title/Summary/Keyword: NIF

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The Role of NifA and PrrA on the Expression of nif Gene in Rhodobacter sphaeroides (Rhodobacter sphaeroides의 nif 유전자의 발현에 대한 NifA와 PrrA의 작용)

  • Son, Myung-Hwa;Kim, Min-Ju;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.21 no.9
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    • pp.1139-1147
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    • 2012
  • To find out the growth conditions for the maximum activity of nitrogenase which catalyzes nitrogen fixation in Rhodobacter sphaeroides, the promoter activities of nifA and nifH were analyzed and the results indicated that expression of both nifA and nifH was increased in response to deprivation of both O2 concentration and nitrogen source. The nifA mutant was constructed by deleting the gene to investigate the effect of NifA, the transcriptional regulator, on the nifH and nifA expression in R. sphaeroides. Analysis of expression of nif genes using the nifA::lacZ and nifH::lacZ fusions in the nifA mutant revealed that NifA acts as a positive activator for nifH and an autoregulator in its own expression. The promoter activities of nifA and nifH in the prrA mutant grown under anaerobic and ${NH_4}^+$-free conditions were derepressed, comparing with those of the wild-type grown under the same conditions, indicating that the prrA product acts as a positive regulator in expression of nifA and nifH.

nif-Gene Organization and Nucleotide Sequence of nifV, nifH, D, K and nifE from Frankia Strain FaCl

  • An, Chung-Sun
    • Proceedings of the Zoological Society Korea Conference
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    • 1995.10b
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    • pp.120-120
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    • 1995
  • The total size of the pF AR1, a genomic clone of Frankia FaCI, was estimated to be about 44Kb by summation of the individual fragment length generated by single or double restriction enzymes. Southern hybridization analyses with Azotobacter vinelandii nif-genes as probes and partial sequencing analyses of the subclones revealed that organization of the nif-gene in the FaCI strain was nifV, H, D, K, E, N, X, W, B. The organization of the structural genes for nitrogenase is the same in this Frankia strain as it is in most other nitrogen-fixing prokaryotes but the positioning of the nifV-like gene relative to the nifHDK cluster differs. A consensus nif-promoter-like sequence, found at 5' of nifH, was not detected upstream of the niJV-like gene. nifV-like gene contained a ORF of 1206 NT encoding 401 amino acids. The nucleotide sequence and deduced amino acid sequence of the gene exhibit homology value of 65% and 41% with that from A vinelandii, respectively. The putative Shine-Dargamo sequences were present preceding nitK, nifH, D, K, and nifE, and in nitK gene putative start codon GTG was detected instead of A TG. The nucleotide and amino acid sequence of niIK of FaCI showed 82% and 76% homolgy with those of Frankia HFPCc 13, respectively. Amino acid sequence of niIK showed 69% and 61% homology with those of A vinelandii, Klebsiella pnewnoniae, respectively, while that of nifE 73% and 71%, respecti vely.i vely.

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Molecular Cloning of nifHD from Rhizobium sp. SNU003 (Rhizobium sp. SNU003의 nifHD 클로닝)

  • 강명수;안정선
    • Korean Journal of Microbiology
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    • v.31 no.2
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    • pp.123-128
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    • 1993
  • Genes for dinitrogenase reductase (nifH) and dinitogenase a subunit (nifD) were found to be located on 7.9 kb of EcoRI, 6.5 kb of Sail, 7.3 kb of HindlII and 4.4 kb of Pstl fragments of the genomic blot of Rhizobium sp. SNU003. a symbiotic strain from root nodule of Canavalia lineata. Nine recombinant phage nif-clones were selected from the genomic library constructed by using EMBL-3 BamHI arms of bacteriophage lambda. Among them. Rnif-6 had insert DNA of 15.3 kb. in which 7.6 kb of BamHI!SacI fragment contained nifHD region. Therefore, the 7.6 kb fragment was subcloned into pUC19 and partial restriction map was constructed. As the results, nifH and nifD were found to be located continuously on 4.5 kb of BamHI/BglIl in the genome of Rhizobium sp. SNU003 strain.

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Molecular Cloning of nifH, D from Frankia EuIK1 Strain, A Symbiont of Elaeagnus umbellata Root Nodules (보리수나무 뿌리혹 공생균주인 Frankia EuIK1의 nifH, D클로닝)

  • Kim, Ho-Bang;Kim, Chun-Ho;Song, Seun-Dal;An, Chung-Sun
    • Korean Journal of Microbiology
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    • v.32 no.4
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    • pp.258-263
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    • 1994
  • Genomic Southern hybridization of Frankia EuIKl strain, a nitrogen fixing symbiont of Elaeagnus umbellate root nodules, with nifH,D of K. pneumoniae as a probe, showed that 3.2 Kb and 5.5 Kb of BamHI fragments and 15 Kb PstI fragment were strongly hybridized with the probe, indicating nifH,D are located on these fragments. Using the same probe, one clone(pEuNIF) was isolated from the genomic library constructed into pWE15 cosmid vector by colony hybridization. The 3.2 Kb and 5.5 Kb BamHI fragments of this clone were hybridized with the same probe and this result corresponds to the genomic Southern hybridization data. However, using nifH of Frankia FaCl strain as a probe, only the 3.2 Kb BamHI fragment showed hybridization signal. Amino acid sequence deduced from nucleotide sequence of 3' terminus of the 3.2 Kb and 5' terminus of the 5.5 Kb fragments showed that the former was highly homologous with that of ArI3 nifD from 182nd to 240th amino acids, while the latter was from 241st to 282nd amino acids. These results show that nifH and partial nifD sequences are located on the 3.2 Kb fragment and residual sequences of nifH on the 5.5 Kb fragment which is contiguous to the 3.2 Kb fragment.

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Frankia sp. strain SNU 014201의 nif-H, D, K, 유전자 클로닝

  • 권석윤;강명수;안정선
    • Korean Journal of Microbiology
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    • v.30 no.1
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    • pp.30-36
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    • 1992
  • nif (nitrogen fixation)-H.D, K genes of Frankia sp. SNU 014201. a symbiotic strain isolated from root nodule of Alnus hirsura, were found to be located in the genome on 13.5 kb of EcoRI, 18.0 kb of BamHI, 10.5 kb of BglII and 4.5 kb of KpnI fragments. Using EMBL-3 BamHI arms of bacteriophage lambda. the genomic library was constructed. from which fourteen recombinant phage nif-clones were selected. Among them, Ahnif-I2 had insert DNA of 18 kb, in which 7.9 kb of BamHl fragment contained nif-H, D, K and 3.6 kb of HindlIl/KpnI had nif-H and partial -D. Therefore, the 7.9 kb and 3.6 kb fragments were subcloned and partial restriction maps were constructed. As the results, nif-F1, D.K genes were found to be located continuously on the 6.5 kb of HindII/BamHI and 5.2 kb of SalIIBamHI fragment in the genome of Frankia sp. SNU 014201.

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Isolation of Nif$^{-10}$ -mutants through transposon mutagenesis in enterobacter agglomerans 339 (Enterobacter agglomerans 339에 있어서 transposon umtagenesis를 통한 Nif$^{-10}$ -mutants 분리 동정)

  • 민병환;이호자
    • Korean Journal of Microbiology
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    • v.26 no.1
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    • pp.20-26
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    • 1988
  • Three $NIf^{-}$ -mutants were isolated from Enterbacter agglomerans 339 through the transposon umtagenesis using a RP4-mobilising system for its nif-gene characterization. All mutants hadn't acetylene-reduction ability. Then we confirmed that Tn5 was inserted into all conserved nif-plasmids through the Southern Hybridization.

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Structural Insights and Mechanistic Understanding of Iron-Molybdenum Cofactor Biosynthesis by NifB in Nitrogenase Assembly Process

  • Wonchull Kang
    • Molecules and Cells
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    • v.46 no.12
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    • pp.736-742
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    • 2023
  • NifB, a radical S-adenosylmethionine (SAM) enzyme, is pivotal in the biosynthesis of the iron-molybdenum cofactor (FeMo-co), commonly referred to as the M-cluster. This cofactor, located within the active site of nitrogenase, is essential for the conversion of dinitrogen (N2) to NH3. Recognized as the most intricate metallocluster in nature, FeMo-co biosynthesis involves multiple proteins and a sequence of steps. Of particular significance, NifB directs the fusion of two [Fe4S4] clusters to assemble the 8Fe core, while also incorporating an interstitial carbide. Although NifB has been extensively studied, its molecular mechanisms remain elusive. In this review, we explore recent structural analyses of NifB and provide a comprehensive overview of the established catalytic mechanisms. We propose prospective directions for future research, emphasizing the relevance to biochemistry, agriculture, and environmental science. The goal of this review is to lay a solid foundation for future endeavors aimed at elucidating the atomic details of FeMo-co biosynthesis.

Cloning of nif genes from Enterobacter agglomerans in Escherichia coli. (Enterobacter agglomerans의 질소고정유전자 Cloning)

  • 정건섭;이정기;민태익;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.15 no.2
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    • pp.116-121
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    • 1987
  • In order to cloning of the nif genes of Enterobacter agglomerans NFB-264, the digested total DNA of the strain was ligated to pBR 322 and transformed into E. coli. Through the negative selection and colony hybridization, the transformants were obtained. The recombinant plasmids, pNEL 10 and pNES 20 were extracted from these transformants. It was known from Southern hybridization that pNEL 10 contained the 12 Mdal foreign DNA fragment hybridized with nif Q-X probe and pNES 20 included the 5 Mdal foreign DNA fragment hybridized with nif NE and nif YK probe.

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The Effects of Nitrogen Sources on the Expression of Nif Gene in Klebsiella pneumoniae Nif-Lac Fusants (Klebsiella pneumoniae nif-lac 융합변이주의 질소고정 유전자 발현에 미치는 질소원의 효과)

  • 김성훈;손형진;김창진;민태익
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.20-24
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    • 1985
  • The effects of various nitrogen soruces on the expression of nif gene were investigated using nif-lac fusants of Klebsiella pneumoniae. K. pneumoniae UK 2979 was infected with Mudl lysate prepared by heat induction of K. pneumoniae UK 4482. About 80 nif-lac fusants were isolated and designated as LX series. In the prescence of $NH_4^+,\;{\beta}-galactosidase$ activities on nif-lac fusants were greatly repressed. Amino acids, such as serine, glutamine and asparagine, were found to support the growth of K. pneumoniae M5al quite well, and showed a repressive effect on ${\beta}-galactosidase$ activities of nif-lac fusants LX-9 and LX-22 in NFHM. Glutamic acid, histidine and arginine rendered poor growth but high activities of ${\beta}-galactosidase$. Good cell growth and high enzyme activity were observed when complex nitrogen sources, such as casitone, proteose pepone, were employed. ${\beta}-galactosidase$ activities of LX-9 and LX-22 in nitrogen free minimal medium increased sharply within first 4 hours.

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NIF Application for Korean Natural Language Processing (한국어 자연언어처리의 NIF 적용에 관한 연구)

  • Seo, Jiwoo;Won, Yousung;Kim, Jeongwook;Hahm, YoungGyun;Choi, Key-Sun
    • Annual Conference on Human and Language Technology
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    • 2014.10a
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    • pp.167-172
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    • 2014
  • 본 논문에서는 한국어 자연언어처리 결과물들을 통일된 형식으로 표준화하기 위해서 NIF를 적용한 내용을 다룬다. 한국어 자연언어처리에 NIF 온톨로지를 적용한 이유와 적용과정에서 야기된 문제점들을 논의한다. 한국어 NLP2RDF 구축과정에서 한국어 자연언어처리에 필요한 새로운 클래스와 프로퍼티들을 추가로 정의하여 NIF 온톨로지를 변형 적용하였다.

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