Construction of Binary Vectors for the Rice Transformation Using a Rice Actin Promoter and Replication Origin of pTi12 Isolated from Agrobacterium tumefaciens KU12

Agrobactrium tumefaciens KU12로부터 분리한 pli12의 Replication Origin과 벼의 Actin 유전자 프로모터를 이용한 벼의 Binary Vector 제조

Binary vectors, pBI-ActR1, pBI-ActF1 and pBSH-ActR1, were constructed using pGA642, the replication origin of pTi12 and the rice actin promoter. The sizes of pBI-ActR1, pBI-ActF1 and pBSH-ActR1 were 12.9 kb, 13.2 kb and 11.95 kb, respectively. These vectors containing a rice actin promoter followed by a GUS structural gene could induce stronly the expression of GUS gene in transformed rice cells. Rice explants from 3-4 day old seedlings after germinatin were cocultured with A. tumefaceins harboring pBI-ActR1, pBI-ActF1 or pBSH-ActR1, and then GUS expression in the explants was assayed. Transformation of rice explants by these binary vectors was tissue-specific, such that the meristematic regions of shoot apex, root and hypocotyl were transformed by these binary vectors.