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.
Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Japan.
M
M
Seki
P
P
Carninci
Y
Y
Nishiyama
Y
Y
Hayashizaki
K
K
Shinozaki
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