Journal of the Korean Society of Tobacco Science (한국연초학회지)

The Korean Society of Tobacco Science (한국연초학회)
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Cytotoxicity Test of Pokeweed Antiviral Protein Type I Gene by Using Yeast Expression System

효모 발현 시스템을 이용 PAP(Pokeweed Antiviral Protein) 유전자의 세포독성 연구

  • Published : 2001.12.01
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Abstract

PAP-I cDNA was synthesized from total RNA of Phytolacca americana leaves by RT-PCR, and then subcloned to recombinant vector pBluescript II SK-. Using PCR with primers designed in our laboratory, we could get the 9 deletion mutant PAP-I cDNA fragments. The first of the fragments was deleted by 66bp from immature N-terminal and then the rest were deleted by 90bp sequentially. Sequentially deletion mutant PAP-I cDNAs were inserted to pAc55M, on down-stream of gall promoter. Recombinant pAc55M was transformed to yeast cells, psy1 and the cells were spreaded on SC_urn-/glucose plate media. Colonies on SC_ura-/glucose plate were streaked on the same position of SC_ura-/glucose and SC_ura-/galactose plate, and we selected colonies growing on both plates, which carry non-cytotoxic deleted mutant PAP-I cDNA. We selected 4 deletion mutant PAP-I cDNAs which have not cytotoxicity.

Keywords

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