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http://dx.doi.org/10.5010/JPB.2003.30.4.307

Molecular Cloning of Differentially Expressed Genes in First Trap Leaf of Dionaea muscipula by Fluorescent Differential Display  

Kang, Kwon-Kyoo (Department of Horticulture, Hankyong National University)
Lee, Keun-Hyang (Department of Horticulture, Hankyong National University)
Park, Jin-Heui (Division of Biotechnology and Genetic Engineering, College of Life and Environmental Science, Korea University)
Hong, Kyong-Ei (Department of Horticulture, Hankyong National University)
Publication Information
Journal of Plant Biotechnology / v.30, no.4, 2003 , pp. 307-313 More about this Journal
Abstract
Fluorescent differential display (FDD) is a method for identifying differentially expressed genes in eukaryotic cells. The mRNA FDD technology works by systematic amplification of the 3' terminal regions of mRNAs. This method involve the reverse transcription using anchored primers designed to bind 5'boundary of the poly A tails, followed by polymerase chain reaction (PCR) amplification with additional upstream primers of arbitrary sequences. The amplified cDNA subpopulations are separated by denaturing polyacrylamide electrophoresis. To identify the genes involved in the development of first trap leaf, we applied a FDD method using mRNAs from leaf base, first trap leaf and flower tissue, respectively. We screened several genes that expressed specifically in first trap leaf. Nucleotide sequence analysis of these genes revealed that these were protease inhibitor (PI), myo-inositol-1-phosphate synthase and lipocalin-type prostaglandin D synthase. Northern blot analysis showed that these genes were expressed specifically in first trap leaf (in vivo and in vitro). FDD could prove to be useful for simultaneous scanning of transcripts from multiple cDNA samples and faster selection of differentially expressed transcripts of interest.
Keywords
Polyacrylamide gel electrophoresis; polymerase chain reaction (PCR); first trap leaf specific gene;
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