The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Isolation and Sequencing of the cDNA Encoding ${\beta}-tubulin$ from Pleurotus sajor-caju

여름느타리버섯으로부터 ${\beta}-tubulin$ cDNA의 분리 및 염기서열 결정

  • Kim, Beom-Gi (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • Shin, Pyung-Gyun (Division of Molecular Genetics, National Institute of Agricultural Science and Technology, RDA) ;
  • 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) ;
  • 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) ;
  • Kwon, Suk-Tae (Department of Genetic Engineering, Sung Kyun Kwan University)
  • 김범기 (농업과학기술원 분자유전과) ;
  • 신평균 (농업과학기술원 분자유전과) ;
  • 정미정 (농업과학기술원 분자유전과) ;
  • 박수철 (농업과학기술원 분자유전과) ;
  • 유영복 (농업과학기술원 분자유전과) ;
  • 류진창 (농업과학기술원 분자유전과) ;
  • 권석태 (성균관대학교 유전공학과)
  • Published : 1997.03.30
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Abstract

The cDNA encoding ${\beta}-tubulin$ of Pleurotus sajor-caju was isolated using an internal gene segment probe amplified by polymerase chain reaction (PCR) of genomic DNA and by cDNA library screening. The cDNA was consisted of 1560 nucleotides(nt), including a 5'-untranslation region (UTR) of 27nt, an open reading frame (ORF) of 1341nt, and a 3'-UTR of 191nt. The ORF encoded a protein of 446 amino acids(aa), which shows over 80% homology with ${\beta}-tubulins$ of other filamentous fungi. Southern hybridization analysis showed that there were two isotypes of ${\beta}-tubulin$ genes in P. sajor-caju. Through sequence analysis we found that ${\beta}-tubulin$ had a unusual $Cys^{165}$ residue, which might be a significant factor for the insensitivity of fungi to fungicide benomyl.

여름느타리버섯 균사체의 cDNA library로부터 ${\beta}-tubulin$ 유전자를 분리하여 염기서열을 분석하였다. 분리된 ${\beta}-tubulin$ cDNA유전자는 27nt의 5'-untranslation region과 1341nt의 open reading frame, 191nt의 3'-untranslation region으로 구성되어 있었다. ORF는 445개의 아미노산들로 구성되어 있으며, 동물, 식물, 사상균에서 보고된 ${\beta}-tubulin$과 80% 이상의 상동성을 보였다. 분리된 Myc7tub clone을 사용하여 Southern hybridization한 결과 여름느타리버섯에는 두 가지의 isotype ${\beta}-tubulin$이 존재할 것으로 생각된다.

Keywords

References

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