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Construction of a Plant Transformation-ready Expression cDNA Library for Thellungiella halophila Using Recombination Cloning
Authors:Wan-Song Ni  Zhi-Yong Lei  Xi Chen  David J. Oliver   Cheng-Bin Xiang
Affiliation:( School of Life Sciences, University of Science and Technology of China, Hefei 230027, China;;Department of Genetics, Development and Cell Biology, Iowa State University, Ames, IA 50011, USA;)
Abstract:Salt cress (Thellungiella halophila), a close relative of the model plant Arabidopsis thaliana L., is an extremophile that is adapted to harsh saline environments. To mine salt-tolerance genes from this species, we constructed an entry cDNA library from the salt cress plant treated with salt-stress by using a modified cDNA synthesis and an improved recombinationassisted cDNA library construction method that is completely free of manipulations involving restriction enzymes and DNA ligase. This cDNA library construction procedure is significantly simplified and the quality of the cDNA library is improved. This entry cDNA library was subsequently shuttled into the destination binary vector pCB406 designed for plant transformation and expression via recombination-assisted cloning. The library is plant transformation ready and is used to transform Arabidopsis on a large scale in order to create a large collection of transgenic lines for functional gene mining.
Keywords:cDNA library construction  cDNA synthesis  recombination assisted  transformation.
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