[KSCI] Korea Science Citation Index Service

http://dx.doi.org/10.5657/fas.2006.9.4.135

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)

Publication Information

Fisheries and Aquatic Sciences / v.9, no.4, 2006 , pp. 135-139 More about this Journal

Abstract

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.

Keywords

GUS gene; Particle bombardment; Porphyra yezoensis; Transformation;

Citations & Related Records

Reference

1	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
2	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
3	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 DOI
4	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 DOI
5	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
6	Sunagawa, M. and Y. Magae. 2002. Transformation of the edible mushroom Pleurotus ostreatus by particle bombardment. FEMS Microbiol. Let., 211, 143-146 DOI
7	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 DOI ScienceOn
8	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 DOI ScienceOn
9	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
10	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
11	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 DOI
12	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
13	Halford, N.G. and P.R. Shewry. 2000. Genetically modified crops: methodology, benefits, regulation and public concerns. Br. Med. Bull., 56, 62-73 DOI ScienceOn
14	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 DOI ScienceOn
15	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
16	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 DOI ScienceOn
17	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
18	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 DOI ScienceOn