Rapid construction of a transcription map for a cosmid contig of Arabidopsis thaliana genome using a novel cDNA selection method.
Significant progress has been made on the random sequencing of cDNAs (ESTs) and the genetic and physical mapping of the Arabidopsis thaliana genome. New techniques are now required to identify and map the expressed genes efficiently on A. thaliana chromosomes. A novel method to construct a transcription map of expressed genes or cDNAs in specific regions of the genome using DNA-latex particles has been developed. The region-specific DNA fragments prepared from six cosmid clones that constitute a contig covering the abi1 locus on chromosome 4 were covalently bound to latex particles. The DNA-latex particles were used for the selection of region-specific cDNAs. Sequence analysis of the cDNA clones revealed that ABI1, RPS2, casein kinase 1 (CK1), nucleosome assembly protein I (NAP) cDNAs and T20837 EST, which are situated within the contig near abi1 locus, were selected. These results indicate that the cDNAs in the specific region of the genome were faithfully selected with this method. Sequence analysis also indicated that 11 selected cDNAs were derived from novel genes located near the abi1 locus and that four of the selected cDNAs encode putative proteins that have sequence similarity to cationic peroxidase, phosphatidylserine decarboxylase 2 (PSD2), trans-caffeoyl CoA 3-O-methyltransferase (CCoAMT), and proteasome subunit XC3.
Seki
M
M
Laboratory of Plant Molecular Biology, Tsukuba Life Science Center, Institute of Physical and Chemical Research (RIKEN), Japan.
Hayashida
N
N
Kato
N
N
Yohda
M
M
Shinozaki
K
K
D017360:Arabidopsis / Q000235:genetics*; D002874:Chromosome Mapping / Q000379:methods; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D005819:Genetic Markers; D018745:Genome, Plant; D015698:Genomic Library; D016133:Polymerase Chain Reaction / Q000379:methods; D012641:Selection, Genetic; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D005819:Genetic Markers