Fisheries and Aquatic Sciences

한국수산과학회 (The Korean Society of Fisheries and Aquatic Science)
권호 표지
DOI QR코드

DOI QR Code

Optimization of Parameters for GUS Gene Transformation of Porphyra yezoensis by Particle Bombardment

  • Nam, Bo-Hye (Biotechnology Research Center, National Fisheries Research & Development Institute) ;
  • Park, Jung-Youn (Biotechnology Research Center, National Fisheries Research & Development Institute) ;
  • Jin, Deuk-Hee (Faculty of Marine Bioscience and Technology, Kangnung National University) ;
  • Hong, Yong-Ki (Department of Biotechnology, Pukyong National University)
  • 발행 : 2006.12.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

We optimized the biological and physical parameters for DNA delivery into thalli of the red alga Porphyra yezoensis using a particle bombardment device. The efficiency of transformation was determined using the ${\beta}-glucuronidase$ (GUS) assay. The optimal helium pressure, distance of tungsten particle flight, and ratio of DNA to tungsten particles were $23kgf/cm^2$, 8 cm, and $5{\mu}g/mg$ tungsten, respectively. During bombardment, osmotic treatment with a mixture of 0.6 M mannitol and sorbitol increased the efficiency of GUS transformation. After 2 days, the blue color indicating GUS activity was observed using a histochemical assay.

키워드

참고문헌

  1. Athmaram T.N., G. Bali and K.M. Devaiah. 2006. Integration and expression of Bluetongue VP2 gene in somatic embryos of peanut through particle bombardment method. Vaccine, 24, 2994-3000 https://doi.org/10.1016/j.vaccine.2005.11.019
  2. Charest, P.J., N. Calero, D. Lachance, R.S.S. Datla, L.C. Duchesne and E.W.T. Tsang. 1993. MicroprojectileDNA delivery in conifer species: factors affecting assessment of transient gene expression using the $\beta$- glucuronidase reporter gene. Plant Cell Rep., 12, 189-193
  3. Cho, S.H., Y.S. Chung, S.K. Cho, Y.W. Rim and J.S. Shin. 1999. Particle bombardment-mediated transformation and GFP expression in the moss Physcomitrella patens. Mol. Cells, 9, 14-19
  4. Choi, J.S., J.Y. Cho, L.G. Jin, H.J. Jin and Y.K. Hong. 2002. Procedures for the axenic isolation of conchocelis and monospores from the red seaweed Porphyra yezoensis. J. Appl. Phycol., 14, 115-121 https://doi.org/10.1023/A:1019504203660
  5. Finer J.J., P. Vain, M.W. Jones and M.D. McMullen. 1992. Development of the particle inflow gun for DNA delivery to plant cells. Plant Cell Rep., 11, 323-328
  6. Halford, N.G. and P.R. Shewry. 2000. Genetically modified crops: methodology, benefits, regulation and public concerns. Br. Med. Bull., 56, 62-73 https://doi.org/10.1258/0007142001902978
  7. Hodal, L., A Bochardt, J.E. Nielsen, O. Mattsson and FT Okkels. 1992. Detection, expression and specific elimination of endogenous beta-glucuronidase activity in transgenic and non-transgenic plants. Plant Sci., 87, 115-122 https://doi.org/10.1016/0168-9452(92)90199-V
  8. Hong, Y.K., S.D. Kim, M. Polne-Fuller and A. Gibor. 1995. DNA extraction conditions from Porphyra perforata using LiCI. J. Appl. Phycol., 7, 101-107 https://doi.org/10.1007/BF00693055
  9. Hu, C.Y., P.P. Chee, R.H. Dhesney, F.H. Zhou, P.D. Miller and T O'Brien. 1990. Intrinsic GUS-like activities in seed plants. Plant Cell Rep., 9, 1-5
  10. Huang, X., J.C. Weber, T.K. Hinson, A.C. Mathieson and S.C. Minocha. 1996. Transient expression of the GUS reporter gene in the protoplasts and partially digested cells of Ulva lactuca L. (Chlorophyta). Bot. Mar., 39, 467-474 https://doi.org/10.1515/botm.1996.39.1-6.467
  11. Jefferson, R.A., T.A Kavanagh and M.W. Bevan. 1987. GUS fusion: B-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO, 6, 3901-3907
  12. Kosugi, S., Y. Ohashi, K. Nakajiama and K. Arai. 1990. An improved assay for beta-glucuronidase in transformed cells: methanol almost completely sup-presses a putative endogenous beta-glucuronidase activity. Plant Sci., 70, 133-140 https://doi.org/10.1016/0168-9452(90)90042-M
  13. Kuang, M. and S.J. Wang. 1998. Transient expression of GUS gene in four tissues of rhodophytes using a biolistic particle delivery system. J. Fish. China, 22, 178-181
  14. Lowry, O.H., N.J. Rosebrough, A.L. FaIT and R.J. Randall. 1951. Protein measurement with the Folin phenol reagent. J. BioI. Chem., 193, 265-275
  15. Nam, B.H., H.J. Jin, S.K Kim and Y.K. Hong. 1998. Quantitative viability of seaweed tissues assessed with 2,3,5-triphenyltetrazolium chloride. J. Appl. Phycol., 10, 31-36 https://doi.org/10.1023/A:1008059314639
  16. Plegt, L. and RJ. Bion. 1989. $\beta$-Glucuronidase activity during development of the male gametophyte from transgenic and non-transgenic plants. Mol. Gen. Genet., 216, 321-327 https://doi.org/10.1007/BF00334371
  17. Provasoli, L. 1968. Media and prospects for cultivation of marine algae: In: Cultures and Collections of Algae, Watanabe, A and A. Hattori, eds. Jap. Soc. Plant Physiol., Tokyo, 63-75
  18. Sunagawa, M. and Y. Magae. 2002. Transformation of the edible mushroom Pleurotus ostreatus by particle bombardment. FEMS Microbiol. Let., 211, 143-146 https://doi.org/10.1111/j.1574-6968.2002.tb11216.x