• Korean Journal of Microbiology
• /
• v.40 no.2
• /
• pp.178-182
• /
• 2004

In the vegetative hyphae of Aspergillus nidulans, a zymogenic form of the class I chitin synthase activity was successfully measured by the assay condition for Saccharomyces cerevisiae class I chitin synthase, Chsl. The class I chitin synthase activity of the A. nidulans chsC wild type strain was increased about six-fold by trypsin-pretreatment, but that of the chsC disruption strain revealed no increase. Interestingly enough, level of the class I chitin synthase activity of the chsC disruption strain was almost the same as that of the chsC wild type without trypsin-pretreatment. These results indicated that the A. nidulans ChsC activity could be measured by account-ing the class I chitin synthase activity without the trypsin-pretreatment as an internal control. Consistence to the expression pattern of the chsC revealed by northern blot analysis, the activity of ChsC was increased upon reaching the culture time for acquiring developmental competence. Our results shown here also supported the previous report suggesting the possible involvement of ChsC in vegetative hyphal growth of A. nidulans.

• The Korean Journal of Mycology
• /
• v.25 no.3 s.82
• /
• pp.167-175
• /
• 1997

The PCR fragments of two distinct chitin synthase genes, PdCHSl and PdCHS2, were cloned from Penicillium diversum KCTC 6786. The nucleotide sequences of PdCHSl and PdCHS2 contained uninterrupted open reading frames (ORFs) of 570 bp excluding the primer sequence. The similarity analysis of the deduced amino acid sequences using BLASTP indicated that the possible evolutionary relationship between P. diversum and ascomycetous fungi. Multialignment of the deduced amino acid sequences of PdCHSs using CLASTAL W revealed that the PdCHSs fell into two different classes: PdCHSl into Class I and PdCHS2 into Class II of chitin synthase defined by Bowen et al. (1992). By Southern blot analysis, it was shown that each of the two genes is present as a single copy in the genome of P. diversum KCTC 6786.

• Journal of Microbiology
• /
• v.39 no.1
• /
• pp.74-78
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• v.7 no.2
• /
• pp.157-159
• /
• 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.

• Journal of Microbiology
• /
• v.38 no.4
• /
• pp.230-238
• /
• 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.

• Mycobiology
• /
• v.38 no.2
• /
• pp.102-107
• /
• 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.

• The Korean Journal of Mycology
• /
• v.26 no.3 s.86
• /
• pp.354-360
• /
• 1998

We isolated three amplified DNA fragments from P. sajor-caju by Polmerase chain reaction (PCR) using the chithin synthase specific primers. Since the sequence analysis of the these fragments showed significant homology to the other known chitin synthase gene, we regarded these cloned fragments as PsCHS1, PsCHS2, and PsCHS3 according to their size. The PsCHS3, which showed the highest sequence homology (83% identity in amino acid level with ChsI of Rhizopus oligosporus in conserved region), was selected to see expression pattern of the corresponding gene. The result of RT-PCR using internal primer of the PsCHS3 fragment revealed that PsCHS3 gene was only expressed in cap and mycelium but not in stipe. In order to see whether the PsCHS3 gene was to be induced by wounding, the comparison of the mRNA level of this gene between wounded and unwounded mature cap showed at least two times induction of this gene by wounding treatment.

• Bulletin of the Korean Chemical Society
• /
• v.30 no.12
• /
• pp.3092-3094
• /
• 2009

• Proceedings of the Zoological Society Korea Conference
• /
• 2000.10a
• /
• pp.60.2-61
• /
• 2000

No Abstract, See Full Text

• Korean Journal of Microbiology
• /
• v.38 no.4
• /
• pp.230-230
• /
• 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.