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Modified Suppression Subtractive Hybridization Identifies an AP2-containing Protein Involved in Metal Responses in Physcomitrella patens  

Cho, Sung Hyun (Department of Biology, Washington University)
Hoang, Quoc Truong (School of Life Sciences and Biotechnology, Korea University)
Phee, Jeong Won (School of Life Sciences and Biotechnology, Korea University)
Kim, Yun Young (School of Life Sciences and Biotechnology, Korea University)
Shin, Hyun Young (School of Life Sciences and Biotechnology, Korea University)
Shin, Jeong Sheop (School of Life Sciences and Biotechnology, Korea University)
Publication Information
Molecules and Cells / v.23, no.1, 2007 , pp. 100-107 More about this Journal
Abstract
The moss Physcomitrella patens has two life cycles, filamentous protonema and leafy gametophore. A modified from of suppression subtractive hybridization (SSH), mirror orientation selection (MOS), was applied to screen genes differentially expressed in the P. patens protonema. Using reverse Northern blot analysis, differentially expressed clones were identified. The identified genes were involved mainly in metal binding and detoxification. One of these genes was an AP2 (APETALA2) domain-containing protein (PpACP1), which was highly up-regulated in the protonema. Alignment with other AP2/EREBPs (Ethylene Responsive Element Binding Proteins) revealed significant sequence homology of the deduced amino acid sequence in the AP2/EREBP DNA binding domain. Northern analysis under various stress conditions showed that PpACP1 was induced by ethephon, cadmium, copper, ABA, IAA, and cold. In addition, it was highly expressed in suspension-cultured protonema. We suggest that PpACP1 is involved in responses to metals, and that suspension culture enhance the expression of genes responding to metals.
Keywords
AP2-Containing Protein; Physcomitrella patens; Protonema; Stress-induced; Suppression Subtractive Hybridization; Suspension Culture;
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