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PLANT PHYSIOLOGY , Vol 103, Issue 2 359-370, Copyright © 1993 by American Society of Plant Biologists
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MOLECULAR BIOLOGY AND GENE REGULATION |
Generation of Expressed Sequence Tags of Random Root cDNA Clones of Brassica napus by Single-Run Partial Sequencing
Y. S. Park, J. M. Kwak, O. Y. Kwon, Y. S. Kim, D. S. Lee, M. J. Cho, H. H. Lee and H. G. Nam
Department of Life Sciences, Pohang Institute of Science and Technology, P.O. Box 125, Pohang, Kyungbuk, 790-600, South Korea (Y.S.P., J.M.K., O-Y.K., H.G.N.)
Two hundred thirty-seven expressed sequence tags (ESTs) of Brassica napus
were generated by single-run partial sequencing of 197 random root cDNA
clones. A computer search of these root ESTs revealed that 21 ESTs show
significant similarity to the protein-coding sequences in the existing data
bases, including five stress- or defense-related genes and four clones
related to the genes from other kingdoms. Northern blot analysis of the 10
data base-matched cDNA clones revealed that many of the clones are
expressed most abundantly in root but less abundantly in other organs.
However, two clones were highly root specific. The results show that
generation of the root ESTs by partial sequencing of random cDNA clones
along with the expression analysis is an efficient approach to isolate
genes that are functional in plant root in a large scale. We also discuss
the results of the examination of cDNA libraries and sequencing methods
suitable for this approach.
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