Agrobacterium tumefaciens 공동배양법을 이용한 옥수수 형질전환체 생산

• Published in: Journal of plant biotechnology Vol. 32; no. 2; pp. 91 - 95
• Main Authors: 조미애(Mi Ae Cho), 박윤옥(Yun Ok Park), 김진석(Jin Suck Kim), 박기진(Ki Jin Park), 민황기(Hwang Ki Min), 유장렬(Jang Ryol L
• Format: Journal Article
• Language: Korean
• Published: Korean Society for Plant Biotechnology 2005; (사)한국식물생명공학회; 한국식물생명공학회
• Subjects: 농학; β-glucuroinidase(GUS); transformation; maize; Agrobacterium strains; {\beta}$-glucuroinidase (GUS)
• ISSN: 1229-2818; 1598-6365; 2384-1397

Agrobacterium tumefaciens-mediated immature embryo transformation was used to produce transgenic maize. Immature embryo of Hi II genotype were co-cultivated with strains Agrobacterium tumefaciens (C58C1) containing the binary vectors (pPTN290) carrying with Ubiquitin promoter-GUS gene as reporter gene and NOS promoter-nptll gene conferring resistance to paromomycin as selective agent. Seven embryogenic callus lines transformed showed the resistance in paromomycin antibiotics. Histochemical GUS assay showed that 7 individual lines transformed with the GUS gene were positive response among the transformants. Southern blot analysis revealed that the nptll gene segregated and expressed in their progeny. 옥수수 미숙배배양과 Agrobacterium tumefaciens공동배양법에 의해 형질전환체를 생산하였다. Hi II계통의 미숙배를 Ubiquitin 1 promoter-GUS유전자와 선발마커로서 nptII 유전자로 제작된 pPTN290벡터를 C58C1에 도입한 후 형질전환 균주로 사용하였다. 7개의 paromomycin저항성 배 발생캘러스를 얻었으며, GUS양성반응을 나타내는 7개의 독립적인 식물체를 얻었다. Southern분석법에 의하여 $T_1$세대 식물체로부터 nptII유전자가 안정적으로 도입되어 있음을 확인하였다.