title: Isolation and characterization of a cDNA encoding the cysteine proteinase inhibitor, induced upon flower maturation in carnation using suppression subtractive hybridization.
abstract: The suppression subtractive hybridization (SSH) method was used to isolate differentially expressed genes during carnation flower maturation. Five cDNA clones, designated as carnation flower maturation-induced (CFMI), were verified as flower maturation-induced cDNAs. Sequence analysis of five CFMI (CFMI-5, CFMI-6, CFMI-7, CFMI-9, and CFMI-10) clones revealed that one of the clones, CFMI-5, showed high sequence similarity to the cysteine proteinase inhibitor gene, predicted to be involved in flower maturation. The full length cDNA clone CFMI-5 was 531 nucleotides (nts) long and consisted of an open reading frame of 294 nucleotides, encoding a 98 amino acid protein, 12 nucleotides of 5'-untranslated region and 3'-untranslated region (225 nts) with a poly(A)+ tail. The predicted CFMI-5 amino acid sequence had a conserved sequence Gln-Val-Val-Ala-Gly, which corresponds to the active site of proteinase inhibition. Northern blot analysis revealed tissue-specific expression of CFMI-5 transcripts, as the transcripts were expressed preferentially in petals and styles. A PCR-based cDNA subtraction method, termed suppression subtractive hybridization, was identified as a rapid method to screen differentially expressed genes in a short time.
authors.affiliation: Graduate School of Biotechnology, Korea University, Seoul.
authors.forename: J Y
authors.initials: JY
authors.lastname: Kim
authors.forename: Y S
authors.initials: YS
authors.lastname: Chung
authors.forename: K H
authors.initials: KH
authors.lastname: Paek
authors.forename: Y I
authors.initials: YI
authors.lastname: Park
authors.forename: J K
authors.initials: JK
authors.lastname: Kim
authors.forename: S N
authors.initials: SN
authors.lastname: Yu
authors.forename: B J
authors.initials: BJ
authors.lastname: Oh
authors.forename: J S
authors.initials: JS
authors.lastname: Shin
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015853:Cysteine Proteinase Inhibitors / Q000235:genetics*; D018076:DNA, Complementary / Q000032:analysis; D015723:Gene Library; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D018521:Plant Physiological Phenomena; D010940:Plant Proteins; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014018:Tissue Distribution
chemical_list: C402727:CFMI protein, Dianthus caryophyllus; D015853:Cysteine Proteinase Inhibitors; D018076:DNA, Complementary; D010940:Plant Proteins