Construction of a plant transformation-ready expression cDNA library for Thellungiella halophila using recombination cloning

Ni, W.-Song; Lei, Z.-Yong; Chen, X.; Oliver, D., J.; Xiang, C.-Bin

Journal of Integrative Plant Biology 49(9): 1313-1319

2007


ISSN/ISBN: 1672-9072
DOI: 10.1111/j.1744-7909.2007.00483.x
Accession: 015358210

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Abstract
Salt cress (Thellungielia 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 recombination-assisted 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.

Construction of a plant transformation-ready expression cDNA library for Thellungiella halophila using recombination cloning