J Integr Plant Biol. ›› 2007, Vol. 49 ›› Issue (9): 1313-1319.DOI: 10.1111/j.1744-7909.2007.00483.x

• Stress & Phytochemistry • Previous Articles     Next Articles

Construction of a Plant Transformation-ready Expression cDNA Library for Thellungiella halophila Using Recombination Cloning

Wan-Song Ni, Zhi-Yong Lei, Xi Chen, David J. Oliver and Cheng-Bin Xiang   


Salt cress (Thellungiella halophila), a close relative of the model plant Arabidopsis thaliana L., is an extremophile that is adapted to harsh saline environments. Tomine 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.

Key words: cDNA library construction, cDNA synthesis, recombination assisted, transformation.

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