• 제목/요약/키워드: chitin synthase genes

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Penicillium diversum으로부터 두 chitin synthase 유전자 절편의 분리 (Cloning of Two chitin Synthase Gene Fragments from Penicillium diversum)

  • 조성필;이상근;이동훈;배경숙;박희문;맹필재
    • 한국균학회지
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    • 제25권3호통권82호
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    • pp.167-175
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    • 1997
  • Penicillium diversum KCTC 6786으로부터 두개의 chitin synthase 유전자 절편(PdCHSl과 PdCHS2)을 PCR로 증폭하고 cloning하였다. PdCHSl과 PdCHS2는 primer 서열을 제외하면 각각 570 bp 길이의 연속된 open reading frame (ORF)을 포함하고 있었다. BLASTP를 이용하여 유추된 아미노산 서열의 유사도를 분석한 결과, P. diversum은 자낭균류와 상당한 진화적 유연관계가 있음을 알 수 있었다. 이들 아미노산 서열을 CLASTAL W를 이용하여 분석한 결과, 본 실험에서 분리된 두 유전자 절편은 Bowen 등 (1992)이 제시한 몇 부류 중 서로 다른 부류, 즉, PdCHSl은 Class I에, PdCHS2는 Class II에 각각 속하는 것으로 확인되었다. 또한, Southern blot 분석을 통하여, 각 유전자는 P. diversum KCTC 6786의 genome에 1개씩만 존재함을 알 수 있었다.

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Cell Cycle-dependent Expression of Chitin Synthase Genes in Aspergillus nidulans

  • Park, Bum-Chan;Maeng, Pil-Jae;Park, Hee-Moon
    • Journal of Microbiology
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    • 제39권1호
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    • pp.74-78
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    • 2001
  • The transcription of the chitin synthase genes (chss) was cell cycle-regulated in Aspergillus nidulans and the expression pattern was classified into two groups. Group one, containing chsA and chsC, showed decreasing transcription level upon entry into the S-phase and no further variation during the remainder of the cell cycle. However, group two, containing chsB, chsD, and csmA showed a sharp decrease of mRNA level upon entry into the G2-phase and an increase during the M-phase. Our results suggested that the chss, belonging to same group with the similar expression pattern during the cell cycle are functionally linked and that chsD may play a role in hyphal growth and development in A. nidulans.

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Identification of a Domain in Yeast Chitin Synthase 3 Interacting with Chitin Synthase 4 by Two-Hybrid Analysis

  • Park, Hyun-Sook;Shin-Jung-Choi;Nok-Hyun-Park;Chi-Hwa-Kim;Sung-Uk-Kim
    • Journal of Microbiology and Biotechnology
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    • 제12권6호
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    • pp.943-949
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    • 2002
  • It has been proposed that chitin synthase 3 (CHS3)-nediated chitin synthesis during the vegetative cell cycle is regulated by chitin synthase 4 (CHS4) of Saccharomyces cerevisiae. To investigate direct protein-protein interaction between the coding products of these two genes, a domain of Chs3p that is responsible for interaction with Chs4p was identified, using the yeast two-hybrid system. This domain of 54 amino acids, termed MIRC3-4 (Maximum Interacting Region of Chs3p with Chs4p), is well conserved among CHS3 homologs of various fungi. Some mutations in MIRC3-4 resulted in a decrease in the enzymatic activity and chitin contents. Chs3p carrying those mutations exhibited weak interactions with Chs4p, when assayed by the yeast two-hybrid system. Surprisingly, all the mutants were sensitive to Calcofluor regardless of changes in enzymatic activities or chitin contents. This report deals with a core region in MIRC3-4 that affects the interaction with Chs4p.

Cloning and Phylogenetic Analysis of Chitin Synthase Genes from Tricholoma matsutake

  • Suh, Seok-Jong;Kim, Il-Hyeon;Nam, Ju-Hyun;Ghim, Sa-Youl;Bae, Kyung-Sook;Kim, Jong-Guk
    • Mycobiology
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    • 제29권4호
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    • pp.179-182
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    • 2001
  • Chitin synthases(UDP-N-acetyl-D-glucosamine: chitin 4-$\beta$-N-acetyl-D-glucosaminyl transferase, EC 2.4.1.16) catalyze the synthesis of chitin from UDP-N-acetyl-D-glucosamine. Two zymogenic type of chitin synthase gene(TmCHS1 and TmCHS2) were amplified and its nucleotide sequences were determined. By the amino acid comparison and UPGMA tree grouping, TmChs1 and TmChs2 were classified as class II and class IV chitin synthases respectively. The class II type TmChs1 was grouped with others of Agaricales ectomycorrhizal mushroom. Additionally the phylogenetic tree was well adapted to Hymenomycete previously classified by morphological and physiological characteristics.

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Characterization and Phylogenetic Analysis of Chitin Synthase Genes from the Genera Sporobolomyces and Bensingtonia subrorea

  • Nam, Jin-Sik
    • 환경생물
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    • 제23권4호
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    • pp.335-342
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    • 2005
  • We cloned seven genes encoding chitin synthases (CHSs) by PCR amplification from genomic DNAs of four strains of the genus Sporobolomyces and of Bensingtonia subrosea using degenerated primers based on conserved regions of the CHS genes. Though amino acid sequences of these genes were shown similar as 176 to 189 amino acids except SgCHS2, DNA sequences were different in size, which was due to various introns present in seven fragments. Alignment and phylogenetic analysis of their deduced amino acid sequences together with the reported CHS genes of basidiomycetes separated the sequences into classes I, II and III. This analysis also permitted the classification of isolated CHSs; SgCHS1 belongs to class I, BsCHS1, SaCHS1, SgCHS2, SpgCHS1, and SsCHS1 belong to class II, and BsCHS2 belongs to class III. The deduced amino acid sequences involving in class II that were discovered from five strains were also compared with those of other basidiomycetes by CLUSTAL X program. The bootstrap analysis and phylogenetic tree by neighbor-joining method revealed the taxonomic and evolutionary position for four strains of the genus Sporobolomyces and for Bensingtonia subrosea which agreed with the previous classification. The results clearly showed that CHS fragments could be used as a valuable key for the molecular taxonomic and phylogenetic studies of basidiomycetes.

A Partial Nucleotide Sequence of Chitin Synthase (CHS) Gene from Rice Blast Fungus, Pyricularia oryzae and Its Cloning

  • Hwang, Cher-Won;Park, In-Cheol;Yeh, Wan-Hae;Takagi, Masamchi;Ryu, Jin-Chang
    • Journal of Microbiology and Biotechnology
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    • 제7권2호
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    • pp.157-159
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    • 1997
  • A 340-bp chitin synthase gene(CHS) fragment was cloned from the genomic DNA of Pyricularia oryzae using a PCR process with two primer DNAs corresponding to highly conserved sequences within fungal CHS genes. The entire DNA nucleotide sequences of the cloned DNA fragment were determined and analyzed. The amino acid sequences deduced from the nucleotide sequence of the amplified DNA fragment showed 86% homology to that of the Aspergillus fumigatus CHSE gene (9). Using this PCR-amplified DNA, about 2.3 kb of including the PCR fragment of CHSE gene was cloned from genomic library.

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Role of LAMMER Kinase in Cell Wall Biogenesis during Vegetative Growth of Aspergillus nidulans

  • Choi, Yu Kyung;Kang, Eun-Hye;Park, Hee-Moon
    • Mycobiology
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    • 제42권4호
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    • pp.422-426
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    • 2014
  • Depending on the acquisition of developmental competence, the expression of genes for ${\beta}$-1,3-glucan synthase and chitin synthase was affected in different ways by Aspergillus nidulans LAMMER kinase. LAMMER kinase deletion, ${\Delta}lkhA$, led to decrease in ${\beta}$-1,3-glucan, but increase in chitin content. The ${\Delta}lkhA$ strain was also resistant to nikkomycin Z.

Insight Into Genes Involved in the Production of Extracellular Chitinase in a Biocontrol Bacterium Lysobacter enzymogenes C-3

  • Choi, Hoseong;Kim, Hyun Jung;Lee, Jin Hee;Kim, Ji Soo;Park, Seur Kee;Kim, In Seon;Kim, Young Cheol
    • The Plant Pathology Journal
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    • 제28권4호
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    • pp.439-445
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    • 2012
  • The chitinase producing Lysobacter enzymogenes C-3 has previously been shown to suppress plant pathogens in vitro and in the field, but little is known of the regulation of chitinase production, or its role in antimicrobial activity and biocontrol. In this study, we isolated and characterized chitinase-defective mutants by screening the transposon mutants of L. enzymogenes C-3. These mutations disrupted genes involved in diverse functions: glucose-galactose transpoter (gluP), disulfide bond formation protein B (dsbB), Clp protease (clp), and polyamine synthase (speD). The chitinase production of the SpeD mutant was restored by the addition of exogenous spermidine or spermine to the bacterial cultures. The speD and clp mutants lost in vitro antifungal activities against plant fungal pathogens. However, the gluP and dsbB mutants showed similar antifungal activities to that of the wild-type. The growth of the mutants in nutrient rich conditions containing chitin was similar with that of the wild-type. However, growth of the speD and gluP mutants was defective in chitin minimal medium, but was observed no growth retardation in the clp and dsbB mutant on chitin minimal medium. In this study, we identified the four genes might be involved and play different role in the production of extracellular chitinase and antifungal activity in L. enzymogenes C-3.