• Journal of Microbiology
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• 제35권3호
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• pp.222-227
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• 1997

DNA fragments homologous to chitin synthase gene were amplified from the genomic DNA of Beauveria brongniartii by PCR using degenerate primers. Cloning and sequencing of the PCR-amplified fragments led to the identification of a gene, designated BbCHSl. Comparison of the deduced amino acid sequence of BbCHSl with those of other Euascomycetes revealed that BbCHSl is a gene for class II chitin synthase. The Blastp search of the deduced amino acid sequence of BbCHSl displayed the highest rate of similarity, 95.8%, with CHS2 of Metarhizium unisopliae. Phylogenetic analysis of the amino acid sequences confirmed the taxonomic and evolutionary position of B. brongniartii, which was previously derived by traditional fungal classification based on morphological features.

• Journal of Microbiology
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• 제35권3호
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• pp.159-164
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• 1997

The phylogenetic study of Penicilium chrysogenum was performed based on amino acid sequence comparison of chitin synthase. Phylogenetic trees were constructed with the deduced amino acid sequences of the highly conserved region of chitin synthease gene fragments amplified by PCR. The BlasP similarity searcch and the bootstrap analysis of the deduced amino acid sequences of chitin synthase from P. chrysogenum with those form other fungi showed a close evolutionary relationship of Penicillium to ascomycetous fungi, especially to genus Aspergilus. The result from bootstrap analysis of the deduced amino acid sequences of the Class II chitin synthase from ascomyceteous fungi supported the usefulness of the Class II chitin synthease for phylogenetic study of filamentous fungi.

• 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.

• 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.

• Journal of Microbiology
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• 제38권4호
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• pp.230-238
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• 2000

Class III chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class III chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5'flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• 한국균학회지
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• 제26권3호통권86호
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• pp.354-360
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• 1998

여름 느타리 Pleurotus sajor-caju로부터 Chitin synthase(CHS) gene 특이 primer를 이용한 PCR을 통해 3개의 DNA 단편을 분리하여 cloning하였다. 분리된 DNA 단편들을 기존에 보고된 CHS 유전자들과의 염기서열을 분석한 결과, 이들 DNA 단편들 3개가 모두 CHS 유전자의 단편임을 확인하였고, 또한 이들은 각각 서로 다른 종류의 CHS 유전자들임을 알 수 있었다. 한편, RT-PCR 방법을 이용하여 분리된 유전자의 발현 실험을 실시해본 결과, 이들중 하나인 PsCHS3 유전자는 갓과 균사에서만 발현되는 기관특이 발현 특성을 보였으며, 또한 이 유전자는 상처 처리에 의해 그 발현이 증가되는 것을 확인하였다. 이러한 실험결과로 볼 때 p. sajor-caju의 경우, 다른 균류들의 경우처럼 다양한 기능을 가진 여러 종류(최소 3종류)의 CHS 유전자를 보유하고 있으며, 이들 각각은 다른 기관, 또는 다른 생육 단계에 작용하고 있을 것으로 생각되고, 특히 병 방어 기작에도 관여할 것으로 추측되어진다.

• Mycobiology
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• 제38권2호
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• pp.102-107
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• 2010

We identified a gene for $\beta$-1,3-glucan synthesis (GBG1), a nonessential gene whose disruption alters cell wall synthesis enzyme activities and cell wall composition. This gene was cloned by functional complementation of defects in $\beta$-1,3-glucan synthase activity of the the previously isolated Saccharomyces cerevisiae mutant LP0353, which displays a number of cell wall defects at restrictive temperature. Disruption of the GBG1 gene did not affect cell viability or growth rate, but did cause alterations in cell wall synthesis enzyme activities: reduction of $\beta$-1,3-glucan synthase and chitin synthase III activities as well as increased chitin synthase I and II activities. GBG1 disruption also showed altered cell wall composition as well as susceptibility toward cell wall inhibitors such as Zymolyase, Calcofluor white, and Nikkomycin Z. These results indicate that GBG1 plays a role in cell wall biogenesis in S. cerevisiae.

• 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.

• 미생물학회지
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• 제38권4호
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• pp.230-230
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• 2002

Class Ⅲ chitin synthases in filamentous fungi are important for hyphal growth and differentiation of several filamentous fungi. A genomic clone containing the full gene encoding Chs4, a class Ⅲ chitin synthase in Penicillium chrysogenum, was cloned by PCR screening and colony hybridization from the genomic library. Nucleotide sequence analysis and transcript mapping of chs4 revealed an open reading frame (ORF) that consisted of 5 exons and 4 introns and encoded a putative protein of 915 amino acids. Nucleotide sequence analysis of the 5′flanking region of the ORF revealed a potential TATA box and several binding sites for transcription activators. The putative transcription initiation site at -716 position was identified by primer extension and the expression of the chs4 during the vegetative growth was confirmed by Northern blot analysis. Amino acid sequence analysis of the Chs4 revealed at least 5 transmembrane helices and several sites for past-transnational modifications. Comparison of the amino acid sequence of Chs4 with those of other fungi showed a close relationship between P chrysogenum and genus Aspergillus.

• 한국균학회지
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• 제38권1호
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• pp.29-33
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• 2010

KGD1 유전자는 비허용온도에서 세포벽에 결함을 보이는 Saccharomyces cerevisiae LP0353 균주의 베타-1,3-글루칸 합성 효소의 활성을 회복시키는 유전자로 분리되었다. $\alpha$-ketoglutarate dehydrogenase를 암호화하는 KGD1 유전자의 효모의 세포벽 합성과 연관된 기능을 분석하기 위하여 유전자 파괴를 시도하였다. KGD1돌연변이는 생장속도가 감소하고, 키틴 합성 효소들의 활성이 증가하였으며, 세포벽 구성 당류의 함량에 변화를 보였다. 또한 Calcofluor white과 Nikkomycin Z 등과 같은 세포벽 합성 저해물질에 대해 감수성 변화를 나타냈다. 이러한 결과들은 KGD1이 효모의 세포벽 특히 베타-1,6-글루칸과 키틴의 생합성에 영향을 주고 있음을 시사한다.