Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Identification of Korean Ginseng Cytochrome P450 gene and Its Characterization by Transformation System

고려인삼 유래 Cytochrome P450 유전자의 동정 및 형질전환에 의한 특성검정

  • Shim, Ju-Sun (Korean Ginseng Center for Most Valuable Products & Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Kim, Yu-Jin (Korean Ginseng Center for Most Valuable Products & Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Jung, Seok-Kyu (Korean Ginseng Center for Most Valuable Products & Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Kwon, Woo-Saeng (Korean Ginseng Center for Most Valuable Products & Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Kim, Se-Young (Korean Ginseng Center for Most Valuable Products & Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Yang, Deok-Chun (Korean Ginseng Center for Most Valuable Products & Ginseng Genetic Resource Bank, Kyung Hee University)
  • 심주선 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 김유진 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 정석규 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 권우생 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 김세영 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 양덕춘 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행)
  • Published : 2009.09.30
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Abstract

Triterpenoid saponins were synthesized in Panax ginseng C.A. Meyer via the isoprenoid pathway by cyclization of 2,3-oxidosqualene to give primarily oleanane (beta-amyrin) or dammarane triterpenoid skeletons. The triterpenoids are backbone and undergoes various modifications (oxidation, substitution and glycosylation), mediated by cytochrome P450 (CYP)-dependent monooxygenases, glycosyltransferase and other enzymes. This is likely to be due in part to the complexity of the molecules and the lack of pathway intermediates for biochemical studies. A cDNA clone encoding a putative CYP gene was isolated from flower bud of ginseng and transformed into the plant(Nicotiana tabacum cv. Xanthi) and confirmed by PCR analysis. The CYP gene (PgCYP) contained an open reading frame(ORF) encoding mature protein of 500 amino acids. The expression of PgCYP were investigated in transgenic tobacco by reverse transcriptase-polymerase chain reaction (RT-PCR).

인삼으로부터 뽑은 PgCYP 유전자와 표지유전자인 NPTII 유전자를 함유하고 있는 Agrobacterium tumefaciens GV3101균주를 이용하여 담배 잎절편과 공동배양한 후 MS 기본배지에 kanamycin 100 $\mu$g/ml, cefotaxime 500 $\mu$g/l, BA 2 mg/l와 NAA 0.2 mg/l가 첨가된 선발배지에 치상하여 4주 후 항생제가 첨가된 기본배지에서 발근시켰다. 생존한 선발체의 잎을 이용하여 PCR 반응으로 도입유전자의 삽입여부를 확인하였다. 또한 선발된 형질전환체를 이용하여 RT-PCR을 실시하여 PgCYP 유전자가 담배식물에 안정적으로 도입되어 전사되고 있음을 확인하였다.

Keywords

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  1. Classification and characterization of putative cytochrome P450 genes from Panax ginseng C. A. Meyer vol.76, pp.12, 2011, https://doi.org/10.1134/S000629791112008X