High-efficiency cloning of Arabidopsis full-length cDNA by biotinylated CAP trapper.
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 x 10(6)/9 micrograms 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.
Seki
M
M
Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Japan.
Carninci
P
P
Nishiyama
Y
Y
Hayashizaki
Y
Y
Shinozaki
K
K
D017360:Arabidopsis / Q000235:genetics*; D019921:Biotinylation; D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary; D018744:DNA, Plant; D015698:Genomic Library; D012315:RNA Caps; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics
D018076:DNA, Complementary; D018744:DNA, Plant; D012315:RNA Caps; D012333:RNA, Messenger; D018749:RNA, Plant