The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
권호 표지

Isolation and Characterization of a Chitin Synthase Gene Fragments from Pleurotus sajor-caju

여름느타리의 Chitin synthase 유전자 단편분리 및 발현 특성 분석

  • Jeong, Mi-Jeong (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Park, Soo-Chul (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Kim, Bum-Gi (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Yoo, Young-Bok (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Ryu, Jin-Chang (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA)
  • 정미정 (농업과학기술원 분자유전과) ;
  • 박수철 (농업과학기술원 분자유전과) ;
  • 김범기 (농업과학기술원 분자유전과) ;
  • 유영복 (농업과학기술원 분자유전과) ;
  • 류진창 (농업과학기술원 분자유전과)
  • Published : 1998.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

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.

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.

Keywords

References

  1. Mol Microbiol v.4 Isolation of a chitin synthase gene (CHS 1) from Candida albicans by expression in Sacchromyces cerevisiae Au-Young, J.;Robbins, P.W.
  2. Proc. Natl. Acad. Sci. USA v.89 Classification of fungal chitin synthases Bowen, A.R.;Chen-Wu, J.L.;Momany, M.;Young, R.;Szanlszlo, P.Z.;Robbins, P.W.
  3. Cell v.46 The S. cerevisiae structural gene for chitin synthase not required for chitin synthesis in vivo Bulawa, C.E.;Slater, M.;Cabib, E.;Au-Young, J.;Sburlati, A.;Adair, W.L.J.;Robbins, P.W.
  4. J. Gen. Microbiol. v.138 Chitinase and chitin synthase 1: counterbalancing activites in cell seperation of Sacchromyces cerevisiae Cabib, E.R.;Silverman, S.J.;Shaw, J.A.
  5. J. Cell Biol. v.108 Chitin synthase 1, an auxiliary enzyme for chitin synthasis in Saccharomyces cerevisiae Cabib, E.;Sburlati, A.;Bowers, B.;Silverman, S.J.
  6. Proc. Natl. Acad. Sci. USA v.68 the control of morphogenesis: an enzymatic mechanism for the initiation of septum formation in yeast Cabib, E.;Farkas, V.
  7. Mol Microbiol v.6 Expression of chitin synthase genes during yeast and hyphal growth phases of Candida albicans Chen-Wu, J.L.;Zwicker, J.;Bowen, A.R.;Robbins, P.W.
  8. J. Gen. Microbiol. v.99 Biosynthasis of fungal cell wall. Mechanisms and implications Gooday, G.W.
  9. Biotechniques v.16 no.1 A simplified method for the preparation of fungal genomic DNA for PCR and RAPD analysis Graham, G.C.
  10. Mol. Gen. Genet. v.251 The Aspegillus nidulans genes chsA and chsD encode chitin synthases which have redundant functions in conidia formation Motoyama, T.;Fujiwara, M.;Kojima, N.;Horiuchi, H.;Ohta, A.;Takagi, M.
  11. Biosci. Biotech. Biochem. v.58 Isolation and characterization of two chitin synthase genes of Rhizopus oligosporus Motoyama, T.;Sudoh, M.;Horiuchi, H.;Ohta, A.;Takagi, M.
  12. J. Biol. Chem. v.262 Two chitin synthases in Saccharomyces cerevisiae Orlean, P.
  13. Molecular cloning. A laboratory manual Sambrook, J.;Fritsch, E.F.;Maniatis, T.
  14. Proc. Natl. had. Sci. USA v.74 DNA sequencing with chain terminating inhibitors Sanger, F.;Nicklen, S.;Coulson, A.R.
  15. J. Cell Biol. v.114 The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle Shaw, J.A.;Mol, P.C.;Bower, B.S.;Silverman, J.M.;Valdiviesd, H.;Duran, A.;Cabib, E.
  16. Proc. Natl. Acad. Sci. USA v.85 Chitin synthase 2 is essential for septum formation and cell division in Saccharomyces cerevisae Silverman, S.J.;Sburlti, A.;Slater, M.L.;Cabib, E.
  17. J. Cell Biol. v.114 CALI, a gene required for activity of chitin synthase 3 in Sacchromyces cerevisiae Valdivieso, M.H.;Mol, P.C.;Shaw, J.A.;Cabib, E.;Duran, A.
  18. Biosci. Biotech. Biochem. v.58 Isolation and characterization of two chitin synthase genes from Aspergillus nidulans Yanai, K.;Kojima, N.;Takaya, N.;Horiuchi, H.;Ohta, A.;Takagi, M.
  19. Gene & Dev v.5 Chitin synthase 1 plays a major role in cell wall biogenesis of Neurospora crass Yarden, O.;Yanofsky, C.