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고구마 배양세포에서 Peroxiredoxin cDNA의 분리 및 발현 특성

Molecular Cloning and Characterization of a Peroxiredoxin cDNA from Cell Cultures of Sweetpotato

  • 박수영 (한국생명공학연구원 환경생명공학연구실) ;
  • 류선화 (한국생명공학연구원 환경생명공학연구실) ;
  • 권석윤 (한국생명공학연구원 환경생명공학연구실) ;
  • 김종국 (경북대학교 미생물학과) ;
  • 곽상수 (한국생명공학연구원 환경생명공학연구실)
  • Park, Soo-Young (Laboratory of Environmental Biotechnology, Korea Research Institute of Bioscience Biotechnology (KRIBB)) ;
  • Ryu, Sun-Hwa (Laboratory of Environmental Biotechnology, Korea Research Institute of Bioscience Biotechnology (KRIBB)) ;
  • Kwon, Suk-Yoon (Laboratory of Environmental Biotechnology, Korea Research Institute of Bioscience Biotechnology (KRIBB)) ;
  • Kim, Jong-Guk (Department of Microbiology, Kyungpook National University) ;
  • Kwak, Sang-Soo (Laboratory of Environmental Biotechnology, Korea Research Institute of Bioscience Biotechnology (KRIBB))
  • 발행 : 2003.06.01

초록

Peroxiredoxin(Pix) are large family of peroxidases that reduce alkyl hydroperoxides and hydrogen peroxide. A cDNA clone (referred to as swPrxl) encoding Pix was from a sweetpotato cDNA library constructed from suspension-sultured cells, and its expression was investigated in terms of stress. The swPrxl contained an open reading frame (ORF) encoding mature protein of 193 amino acids with calculated molecular mass of 20.8kDa. The predicted amino acid sequence of swPrxl has two conserved cysteines that are essential resicues for the reduction of peroxides. It showed high amino acid sequence homology ot PixIIF of Arabidopsis (77%) and putative Prx of rice(72%). RNA gel-blot analysis showed that swPrxl gene was expressed dominantly in leave among intact tissues, and also highly detect in suspension-cultured cells. Interestingly, the level of swPrxl transcripts was almost the same regardless of the growth stage in suspension culture. Furthermore, the transcription level of swPrxl gene was not significantly changed in response to various stress treatments such as wounding, extreme temperature and stress-related chemicals RT-PCR analyses.

키워드

참고문헌

  1. Allen RD, Webb RP, Schake SL (1997) Use of transgenic plants to study antioxidants defenses. Free Rad BioI Med 23: 473-479 https://doi.org/10.1016/S0891-5849(97)00107-X
  2. Altschul SF, Gish W,Miller W,Myers EW, Upman DJ (1990) Basic local alignment search tool. J Mol Biol 215: 403-410 https://doi.org/10.1016/S0022-2836(05)80360-2
  3. Asada K (1992) Ascorbate peroxidase a hydrogen peroxidase-scavenging enzyme in plants. Physiol Plant 85: 235-241 https://doi.org/10.1111/j.1399-3054.1992.tb04728.x
  4. BaierM,Dietz KJ (1997) The plant 2-Cys peroxiredoxin BAS 1 is a nuclear encoded chloroplast protein. Its expresslcnal regulation, phylogenetic origin and implicatirns for its specific physiologtcal functions in plants. Plant J 12: 179-190 https://doi.org/10.1046/j.1365-313X.1997.12010179.x
  5. Baier M, Dietz KJ (1999) Protective function of chloroplast 2-cysteine peroxiredoxin in photosynthesis: evidence from transgenic Arabidopsis. Plant Physiol 119: 1407-1414 https://doi.org/10.1104/pp.119.4.1407
  6. Chol YO, Cheong NE, Lee KO, Jung BG, HongCH, Jerng JH, Chi YH, Kim K, Cho MJ, Lee SY (1999) Cloning and expression of a new isotype of the peroxiredoxin gene of chinesecabbage and its cam parison to 2Cys-peroxiredoxin isolated fran the same plant. Biochem Biophys Res Commun 258: 768-771 https://doi.org/10.1006/bbrc.1999.0714
  7. Cheong NE, Choi YO, Lee KO, Kim WY, Jung BG, Chi YH, Jeong JS, Kim K,Cho MJ, Lee SY (1999)Molocularcloning, expresskn, and functional characterization of a 2-Cys peroxiredoxin in Chinese cabbage. Plant Mol BioI 40: 825-834 https://doi.org/10.1023/A:1006271823973
  8. Choi SM, Lee OS, Kwon SY, Kwak SS, Yu DY, LeeHS (2003) High expression of a human lactoferrin in transgenic tobacco cell cultures. Biotech Lett 25: 213-218 https://doi.org/10.1023/A:1022341917735
  9. Clark CA, Moyer JW (1988) Compendium of sweet potato diseases. APS Press, St. Paul. pp 74
  10. Dellaporta SL, Wood J, Hicks JB (1983) Maize DNA minipreps. Maize Genet Coop Newslett 57: 26-29
  11. Dietz KJ, Horling F, Konig J, Baier M (2002) The function of the chloroplast 2-cysteine peroxiredoxin in peroxide detoxification and its regulation. J Exp Bot 53: 1321-1329 https://doi.org/10.1093/jexbot/53.372.1321
  12. Horling F, BaierM,Dietz, KJ (2001) Redox-regulation of expressirn of peroxide-detoxyfying chloroplast 2- Cysteine peroxiredoxin in liverwort Riccia fluitans. Planta 214: 304-313 https://doi.org/10.1007/s004250100623
  13. Horling F, Konig J, Dietz KJ (2002) Type II peroxiredoxin C, a member of the peroxiredoxin family of Arabidopsis thaliana: its expression and activity in comparison with other perodredoxins. Plant Physiol Biochem 40: 491-499 https://doi.org/10.1016/S0981-9428(02)01396-7
  14. Horling F, Lamkemeyer P, Konig J, Finkemeier I, Kandlbinder A, Baier M, Dietz KJ (2003) Divergent light-, ascorbate-, and codative stress-dependent regulation of expression of the peroxiredoxingene family in Arabidopsis. Plant Physiol 131: 317-325 https://doi.org/10.1104/pp.010017
  15. Huh GH, Lee SJ, Bae YS,Liu JR, Kwak SS (1997) Molocular cloning and characterization of cDNAs for anicnic and neutral peroxidase from suspension-cultured cells of sweetpotato and their differential expression in response to stress. Mol Gen Genet 255: 382-391 https://doi.org/10.1007/s004380050510
  16. Kim KY, Huh GH, Lee HS,Kwon SY, HurY, Kwak SS (1999) Molecular characterization of two anionic peroxidase cDNAs isolated from suspension cultures of sweetpotato. Mol Gen Genet 261: 941-947 https://doi.org/10.1007/s004380051041
  17. Kim KY, Kwon SY, Lee HS, Hur Y, Bang JW, Kwak SS (2003) A novel oxidative stress-inducible peroxidase promoter from sweet potato: molecular cloning and characterization in transgenic tobacco plantsand cultured cells. Plant Mol Biol 51: 831-838 https://doi.org/10.1023/A:1023045218815
  18. Kim SK, Kwak SS, Jung KH, Min SR, Park IH, Liu JR (1994) Selection of plant cell lines for high yields of peroxidase. J Biochem Mol Biol 27: 132-137
  19. Kwak SS, Kim SK,Lee MS, Jungg KH, Park IH, Liu JR (1995) Acidic peroxldases from suspension-cultures of sweetpotato. Phytochemistry 39: 981-984 https://doi.org/10.1016/0031-9422(95)00098-R
  20. Kwon SY, Jerng YJ, Lee HS, Kim JS, Cho KY, Allen RD, Kwak SS (2002) Enhanced tolerance oftransgenictobacco plantsexpressing both superoxide dismutase and ascorbate peroxidase in chloroplasts against methyl viologen-mediated oxidative stress. Plant Cell Environ 25: 873-882 https://doi.org/10.1046/j.1365-3040.2002.00870.x
  21. Lee KO, Jang HH, Jung, BG, Chi YH, Lee JY, Choi YO, Lee JR, Lim CO, Cho MJ, Lee SY (2000) Rice 1Cys-peroxiredoxin overexpressed in transgenictobacco does not maintaindormancybut enhances antioxidant activity. FEBS Lett 486: 103-106 https://doi.org/10.1016/S0014-5793(00)02230-4
  22. Linsmaier EM, Skoog F (1965) Organc growth factor requirements of tobacco tissue cultures. Physiol Plant 18: 100-127 https://doi.org/10.1111/j.1399-3054.1965.tb06874.x
  23. Morgan RW, Christman MF, Jacobson FS, Stortz G, Ames BN (1985) Hydrogen peroxide induced proteins in Salmonelia typhimurium overlap with heat-shock and other stress proteins. Proc Natl Aacd Sci83: 375-379
  24. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473-497 https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  25. Murata N, Ishizaki N, Hayashi H, Tasaka Y, Nishida I (1992) Genetically engineered alteration in the chilling sensitivity of plants. Nature 356: 710-713 https://doi.org/10.1038/356710a0
  26. Naqvi SM, Park KS, Yi ,SY, Lee HW, Bok SH, Chol DI (1998) A glycine-rich RNA-binding protein gene is differentially expressed durlnq acute hypersensitive response following tobacco mosac virus infection in tobacco. Plant Mol Biol 37: 571-576 https://doi.org/10.1023/A:1006031316476
  27. Park SY, Ryu SH, Kwon SY, Lee HS, Kim JG, Kwak SS (2003) Differential expression of six novel peroxidase cDNAs derived from sweetpotato cell cultures in response to stress. Mol Gen Genomics (in press)
  28. Rao MV, Paliyath G, Ormrod DP, Murr DP, Watkins CB (1997) lnnence of salicylic add on $H_20_2$ production, oxidative stress and$H_20_2$ metabolizing enzymes. Plant Physiol 115: 137-149 https://doi.org/10.1104/pp.115.1.137
  29. Stacy RAP, MlU1the E, Steinum T, Sharma B, Aalen RB (1996) A peroxiredoxin antioxidant is encoded by a dormancy-related gene, Per1, expressed during late development in the aleurone and embryoofbarleygrains. Plant Mol Biol 31: 1205-1216 https://doi.org/10.1007/BF00040837