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Construction of a Plant Transformation‐ready Expression cDNA Library for Thellungiella halophila Using Recombination Cloning
Author(s) -
Ni WanSong,
Lei ZhiYong,
Chen Xi,
Oliver David J.,
Xiang ChengBin
Publication year - 2007
Publication title -
journal of integrative plant biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.734
H-Index - 83
eISSN - 1744-7909
pISSN - 1672-9072
DOI - 10.1111/j.1744-7909.2007.00483.x
Subject(s) - cdna library , complementary dna , cloning (programming) , arabidopsis , transformation (genetics) , biology , library , arabidopsis thaliana , insert (composites) , genomic library , molecular cloning , microbiology and biotechnology , gene , genetics , computer science , peptide sequence , mechanical engineering , 16s ribosomal rna , mutant , engineering , programming language
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 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.