[Establishment of promoter trapping system mediated by activator/dissociation (beta-glucuronidase) construction in rice].
The coding region of Bar gene, the left border of Ds element, the coding region of GUS gene, the transposase of Ac element, the right border of Ds element and the promoter of Ubi gene were inserted into the T-DNA region of vector pCAMBIA1300 in turn to construct plasmid p13B. The orientations of the ubiquitons' promoter, Ac transposase and Bar are identical but opposite to that of the GUS gene (Fig.1). The plasmid p13B was then introduced into the calli of Oryza sativa subsp. japonica cv. Zhonghua 11 by Agrobacterium tumefaciens-mediated transforming to trap genes in rice. Eighteen independent transgenic lines were obtained and propagated. T(2) generations of 18 independent transgenic lines were screening by herbicide (Basta) (Fig.2) and the herbicide-resistant plants obtained were analyzed by PCR (Fig.3). Ds element transposed in an inheritable manner was found in 37 plants, in which 5 plants showed GUS activity (Fig.4).
Wang
Ai-Min
AM
Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200032, China.
Chen
Shi-Yan
SY
Shen
Ge-Zhi
GZ
Wang
Xin-Qi
XQ
Ju
Dan-Hua
DH
Wang
Zhong-Lin
ZL
Wang
Zong-Yang
ZY
Cai
Xiu-Ling
XL
D016960:Agrobacterium tumefaciens / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000235:genetics* / Q000378:metabolism; D012275:Oryza / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D014170:Transformation, Genetic
D010940:Plant Proteins; D005966:Glucuronidase