High-efficiency cloning of
Arabidopsis
full-length cDNA by biotinylated CAP trapper |
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| Authors: | Motoaki Seki Piero Carninci Yoko Nishiyama Yoshihide Hayashizaki Kazuo Shinozaki |
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| Institution: | ;Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), 3–1–1 Koyadai, Tsukuba 305–0074, Japan, ;Genome Science Laboratory, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), 3–1–1 Koyadai, Tsukuba 305–0074, Japan, and ;CREST, Japan Science and Technology Corporation (JST), Kawaguchi-Center building, 4–1–8 Hon-cho, Kawaguchi, Saitama 332, Japan |
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| Abstract: | Full-length cDNAs are essential for functional analysis of plant genes. We constructed high-content, full-length cDNA libraries from Arabidopsis thaliana plants based on chemical introduction of a biotin group into the diol residue of the CAP structure of eukaryotic mRNA, followed by RNase I treatment, to select full-length cDNA. More than 90% of the total clones obtained were of full length; recombinant clones were obtained with high efficiency (2.2 × 106/9 μg starting mRNA). Sequence analysis of 111 randomly picked clones indicated that 32 isolated cDNA groups were derived from novel genes in the A. thaliana genome. |
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