Korean Journal of Plant Tissue Culture (식물조직배양학회지)

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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The Introduction of Proteinase Inhibitor II (PI-II) Gene into Flowering Cabbage, Brassica oleracea var. acephala DC.

꽃양배추로의 Proteinase Inhibitor II ( PI-II ) 유전자 도입

  • Published : 1998.01.01
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Abstract

Hypocotyl explants of flowering cabbage were precultured on MS medium without kanamycin and then cocultured with Agrobacterium tumefaciens LBA4404;;pGA875 harboring insect resistantce proteinase inhibitor II(PI-II) gene in MS liquid medium adjusted pH 5.5 for 72hr. These explants were transferred to MS medium containing 20 mg/L kanamycin, 500 mg/L carbenicillin, and 1 mg/L BA. The explants were subsequently subcultured every 2 weeks. After 4 weeks of subculture, kanamycin-resistant shoots were obtained from selection medium. Leaves of putative transformants survived on MS selection medium containing 30 mg/L kanamycin. Incoporation of the PI-II gene into flowering cabbage was confirmed by PCR analysis of genomic DNA. Southern blot analysis showed that ECL-labeled probe for PI-II gene was hybridized to the expected amplified genomic DNA fragment of about 500 by from transgenic flowering cabbage.

꽃양배추의 하배축 조직을 proteinase inhibitor II 유전자가 도입된 Agrobacterium tumefaciens LBA 4404와 2일간 pH 5.5로 조절된 MS 액체배지에서 공동배양후 carbenicillin 500mg/L kanamycin 20mg/L와 BA 1mg/L가 함유된 MS 재분화배지에 옮겼다. 이들 조직을 매 2주마다 계대배양하였으며 약 4주후에 kanamycin 저항성 개체를 얻었다. 형질전환된 것으로 추정되는 식물체는 kanamycin 30mg/L가 함유된 선발배지에서 생존하였다. PCR 분석결과, PI-II 유전자가 형질전환체의 게놈상에 삽입되어 있음을 확인하였다. 형질전환체의 Southern blot 분석을 통하여 ECL-labelling된 PI-II 유전자와 동일한 것으로 판단되는 약 500bp 위치에서 밴드를 확인할 수 있었다.

Keywords

References

  1. Transgenic Plants v.Ⅰ Production of Bacillus thuringiesis insectidal proteins in plants. Barton KA;Miller MJ;Kung S(ed.);Wu R(ed.)
  2. Theor Appl Genet v.76 Genotype-independent leaf disc transformation of potato(Solanum Tuberosum) using Agrobacterim tumefaciens. De Block M
  3. Plant Physiol v.91 Transformation of Brassica napus and Brassia oleracea using Agrobacterium tumefaciens and the expression of the bar and neo genes in the transgenic plants. De Block M;Brouwer DD;Tenning P
  4. Bio/Technology v.7 Field performance of trangenic tomato plants expressing the Bacillus thuringiensis var. kurtaki insect control protein. Delanny X;Lavalle BJ;Proksch RK;Fuchs RL;Slims SR;Greenplate JT;Marroone PG;Dodson RB;Augustine JJ:Layton JG;Fischhoff DA
  5. Plant Cell Reports v.9 The effects of acetosyringone and pH on Agrobacterium-mediated transformation vary according to plant species. Godwin I;Todd G;Ford-Lloyd B;Newbury HJ
  6. Nature v.328 Virus restance in transgenic plants that express cucumber mosaic satellite RNA. Harrison BD;Mary MA;Baulcombe DC
  7. Nature v.300 A nobel machanism of insect resistance engineered into tobacco. Hilder VA;Gatehouse AMR;Sheerman SE;Barker RF;Boulter D
  8. Asian Seed v.3 no.5 Progress of genetic engineering in field and horticulture crops. James C
  9. Proc Natl Acad Sci USA v.86 Expression of Proteinase inhibitor Ⅰ and Ⅱ in transgenic tobaco plant ; Effects on natural defense against Manduca sexta larvae. Johnson R;Narvaez J;An G;Ryan C
  10. Theor Appl Genet v.80 Transformation of Brassica napus canola cultivars with Arabidopsis thaliana acetohydroxyacide synthase genes and analysis of herbicide resistance. Miki BL;Labbe H;Hattori J;Ouellet T;Gabard J;Sunohara G;Charest PJ;Iyer VN
  11. Plant Cell Reports v.8 High efficiency transformation of Brassica napus using Agrobacterium vectors. Moloney MM:Walker JM;Sharma KK
  12. Nucl Acids Res v.8 Rapid isolation of high molecular weight plant DNA. Murray MG;Thompson WF
  13. Plant Cell Reports v.12 Variation amongst Brassica juncea cultivars for regeneration from hypocotyl explants and optimization of conditions for Agrobacterium-mediated genetic transformation. Pental D;Pradhan AK;Sodhi YS;Mukhopadhyay A
  14. Plant Cell Rep v.6 Stimulation of shoot regeneration in Triticum aestivum and Nicotiana plumbagenifolid Viv. tissue cultures using the ethylene inhibitor AgNO₃ Purnhauser L;Medgyesy P;Czako M;Dix PJ;Marton L
  15. Theor Appl Genet v.75 Transformation of Brassica napus L. using Agrobacterium tumefaciens ; developmentally regulated expression of a reintroduced napin gene. Radke SE;Andrews BM;Moloney MM;Crouch ML;Kridl JC;Knauf VC
  16. Science v.233 Engineering herbicide tolerance in transgenic plants. Shah DM;Horsh RB;Klee HH;Kishore GM;Winter JA;Tumer NE;Hiromaka CM;Sanders PR;Gusser CS;Aykent S;Siegal NR;Roger S;Franley RT
  17. J Mol Biol v.98 Detection of specific sequences among DNA fragments separated by gel electrophoresis. Southern E
  18. Japan J Bot v.7 Genome analysis in Brassica with special reference to the experinental formation of B. napus and pecurlar mode of fertization. UN