Cloning of Two chitin Synthase Gene Fragments from Penicillium diversum

Penicillium diversum으로부터 두 chitin synthase 유전자 절편의 분리

  • Cho, Seong-Pil (Department of Microbiology, Chungnam National University) ;
  • Lee, Sang-Keun (Department of Microbiology, Chungnam National University) ;
  • Lee, Dong-Hun (Culture Collection Program, Genetic Resources Center, KRIBB) ;
  • Bae, Kyung-Sook (Culture Collection Program, Genetic Resources Center, KRIBB) ;
  • Park, Hee-Moon (Department of Microbiology, Chungnam National University) ;
  • Maeng, Pil-Jae (Department of Microbiology, Chungnam National University)
  • 조성필 (충남대학교 자연과학대학 미생물학과) ;
  • 이상근 (충남대학교 자연과학대학 미생물학과) ;
  • 이동훈 (생명공학연구소 유전자원센터) ;
  • 배경숙 (생명공학연구소 유전자원센터) ;
  • 박희문 (충남대학교 자연과학대학 미생물학과) ;
  • 맹필재 (충남대학교 자연과학대학 미생물학과)
  • Published : 1997.09.30

Abstract

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.

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.

Keywords

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