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Random sequencing of cDNAs and identification of mRNAs

Published online by Cambridge University Press:  20 January 2017

James V. Anderson  and
David P. Horvath
David P. Horvath
Affiliation:
U.S. Department of Agriculture, Agricultural Research Service, Biosciences Research Laboratory, Plant Science Research, 1605 Albrecht Boulevard, P.O. Box 5674, Fargo, ND 58105
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Abstract

As a first step toward developing a genomics-based research program to study growth and development of underground adventitious shoot buds of leafy spurge, we initiated a leafy spurge expressed sequence tag (EST) database. From the approximately 2,000 clones randomly isolated from a cDNA library made from a population containing growth-induced underground adventitious shoot buds, we have obtained ESTs for 1,105 cDNAs. Approximately 29% of the leafy spurge EST database consists of expressed genes of unknown identity (hypothetical proteins), and 10% represents ribosomal proteins. The remaining 60% of the database is composed of expressed genes that show BLASTX sequence identity scores of ≥ 80 with known GenBank accessions. Clones showing sequence identity to a Histone H3, a gibberellic acid-responsive gene, Tubulin, and a light-harvesting chlorophyll a/b-binding protein were shown to be differentially expressed in underground adventitious shoot buds of leafy spurge after breaking of dormancy. RNA encoding a putative cyclin-dependent protein kinase (CDK)-activating kinase, a gene associated with cell division, and Scarecrow-like 7, a gene involved in GA signaling, were present at similar levels in dormant and growth-induced underground adventitious shoot buds. These data show how even a small EST database can be used to develop a genomics-based research program that will help us identify genes responsive to or involved in the mechanisms controlling underground adventitious shoot bud growth and development.

Keywords

Amp, ampicillinCAK, CDK-activating protein kinaseCDK, cyclin-dependent protein kinaseEST, expressed sequence tagLB, Luria–Bertani brothLhcb, light-harvesting chlorophyll a/b-binding proteinNCBI, National Center for Biotechnology InformationSSC, sodium chloride sodium citrate solutionCell cycledormancygenomicsperennial weedssignal transduction
Type
Research Article
Information
Weed Science , Volume 49 , Issue 5 , October 2001 , pp. 590 - 597
Copyright
Copyright © Weed Science Society of America 

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