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A mini binary vector series for plant transformation
Authors:Xiang  Chengbin  Han  Peng  Lutziger  Isabelle  Wang  Kan  Oliver  David J
Institution:(1) Department of Botany, Iowa State University, 353 Bessey Hall, Ames, IA 50011, USA;(2) Department of Plant Transformation Facility, Iowa State University, Ames, IA 50011, USA
Abstract:A streamlined mini binary vector was constructed that is less than 1/2 the size of the pBIN19 backbone (3.5 kb). This was accomplished by eliminating over 5 kb of non-T-DNA sequences from the pBIN19 vector. The vector still retains all the essential elements required for a binary vector. These include a RK2 replication origin, the nptIII gene conferring kanamycin resistance in bacteria, both the right and left T-DNA borders, and a multiple cloning site (MCS) in between the T-DNA borders to facilitate cloning. Due to the reduced size, more unique restriction sites are available in the MCS, thus allowing more versatile cloning. Since the traF region was not included, it is not possible to mobilize this binary vector into Agrobacterium by triparental mating. This problem can be easily resolved by direct transformation. The mini binary vector has been demonstrated to successfully transform Arabidopsis plants. Based on this mini binary vector, a series of binary vectors were constructed for plant transformation.
Keywords:Agrobacterium tumefaciens  Arabidopsis thaliana  binary vector  T-DNA  transformation
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