생명과학회지 (Journal of Life Science)

  • 제15권6호
  • /
  • Pages.1034-1036
  • /
  • 2005
  • /
  • 1225-9918(pISSN)
  • /
  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
권호 표지
DOI QR코드

DOI QR Code

Sodium hypochlorite처리에 따른 배추종자의 Agrobacterium이용 형질전환 증대

Sodium Hypochlorite Solution As a Chemical Wounding Agent for Improving Agrobacterium-mediated Chinese Cabbage Seed Transformation

  • 신동일 (대구가톨릭대학교생명공학과) ;
  • 박희성 (대구가톨릭대학교생명공학과)
  • Shin Dong-Il (Department of Biotechnology, Catholic University) ;
  • Park Hee-Sung (Department of Biotechnology, Catholic University)
  • 발행 : 2005.12.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

배추 종자 및 유묘에 대하여 GUS발현 또는 hepatitis B surface antigen (HBsAg)발현 벡터를 지니는 Agrobacterium tumefaciens LBA4404 세포를 이용하여 진공침윤(agroinfiltration)에 의한 형질전환을 시도하였다. 특히 ELISA를 이용한 HBsAg발현의 정량적 분석에서 agroinfiltration방법은 형질전환효율이 매우 저조하게 나타났다. 그러나 차아염소산나트륨 용액을 발아 전 또는 발아 중인 배추종자에 처리한 후 agroinfiltration을 실시한 경우 형질전환 효율이 $2\~5$배 증가하였다. 따라서 차아염소산나트륨 등의 화학연마제에 의한 종자의 상처발생이 Agrobacterium의 감염을 용이하게 함으로써 배추유묘에서의 일시유전자발현을 증대시키는 것으로 제안되고있다.

Chinese cabbage (Brassica campestris ssp. napus var. pekinensis Makino) seeds/seedlings were transformed via vacuum-infiltration with recombinant Agrobacterium tumefaciens LBA4404 cells. The agroinfiltration method was determined to be unsuccessful for Chinese cabbage transformation during the analysis of hepatitis B surface antigen expression by ELISA. However, treatment of sodium hypochlorite solution, prior to agroinfiltration, to pregerminated or germinating 1 day- or 2 days-old seeds was proven effectively to enhance transformation efficiency, suggesting that chemical wounding caused by sodium hypochlorite reaction might facilitate Agrobacterium infection and, therefore, transient gene expression in Chinese cabbage sprouts.

키워드

참고문헌

  1. Bechtold, N., J. Ellis and G. Pelletier. 1993. In planta Agrobacterium mediated gene transfer by infiltration of adult Arabidopsis thaliana plants. CR. Acad. Sci. Paris 316, 1194-1199
  2. Cho, H. S., J. Cao, J. P. Ren and E. D. Earle. 2001. Control of lepidopteran insect pests in transgenic Chinese cabbage transformed with synthetic Bacillus thuringiensis cry1C gene. Plant Cell Rep. 20, 1-7 https://doi.org/10.1007/s002990000278
  3. Cho, Y. N., S. Y. Park, T. K. Noh, M. J. Song, Y. S. Park and B. W. Min. 2003. Transformation of Chinese cabbage with L-gluco-$\gamma$-lactone oxidase (GLOase)-encoding gene using Agrobacterium tumefaciens. Kor. J. Hort. Sci. 21, 9-13
  4. Christey, M. C., B. K. Sinclair, R. H. Braun and L. Wyke. 1997. Regeneration of transgenic vegetable Brassicas (Brassica oleracea and B. campestris) via Ri-mediated transformation. Plant Cell Rep. 16, 587-593 https://doi.org/10.1007/BF01275497
  5. Doran, P. M. 2000. Foreign protein production in plant tissue cultures. Curr. Opin. Biotech. 11, 199-204 https://doi.org/10.1016/S0958-1669(00)00086-0
  6. Fisher, R., C. Vaquero-Martin, M. Sack, J. Drossard, N. Emans and U. Commandeur. 1999. Towards molecular farming in the future: transient protein expression in plants. Biotech. Appl. Biochem. 30, 113-116
  7. Giddings, G. 2001. Transgenic plants as protein factories. Curr. Opin. Biotech. 12, 450-454 https://doi.org/10.1016/S0958-1669(00)00244-5
  8. Jefferson, R. A. 1987. Assaying chimeric genes in plants: the gus gene fusion system. Plant Mol. Biol. Rep. 5, 387-405 https://doi.org/10.1007/BF02667740
  9. Jefferson, R. A., T. A. Kavanagh and M. W. Bevan. 1987. GUS fusions: $\beta$-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6, 3901-3907
  10. Joung, Y. H., J. W. Youm, J. H. Jeon, B. C. Lee, C. J. Ryu, H. J. Hong, H. C. Kim, H. Joung and H. S. Kim. 2004. Expression of the hepatitis B surface S and preS2 antigens in tubers of Solanum tuberosum. Plant Cell Rep. 22, 925-930
  11. Sojikul, P., N. Buehner and H. S. Mason. 2003. A plant signal peptide-hepatitis B surface antigen fusion protein with enhanced stability and immunogenecity expressed in plant cells. Proc. Natl. Acad. Sci. USA 100, 2209-2214
  12. Sunil Kumar, G. B., T. R. Ganapathi, C. J. Revathi, K. S. N. Prasad and V. A. Bapat. 2003. Expression of hepatitis B virus surface antigen in tobacco cell suspension cultures. Protein Exrp. Purif. 32, 10-17 https://doi.org/10.1016/j.pep.2003.07.004