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KMID : 0903519960390020112
Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1996 Volume.39 No. 2 p.112 ~ p.117
Molecular Cloning and Nucleotide Sequencing of a DNA Clone Encoding Arginine Decarboxylase in Rice(Oryza sativa L .)




Abstract
Arginine decarboxylase (ADC) is the first enzyme in one of the two pathways of diamine putrescine biosynthesis in plants. The genes encoding ADC have previously been cloned from Escherichia coli, oat and tomato genome. Two degenerate oligonucleotides (17-mer) corresponding to two conserved regions of ADC were used as primers in polymerase chain reaction of rice (Oryza sativa L.) genomic DNA, and an approximately 1.0 kbp fragment was obtained. This amplified PCR product showed an open reading frame which contains 1,022 by of nucleotide sequences. This PCR product was cloned into pGEM-originated T vector anti the short 500 by Pst1 digested fragment was subcloned into pGEM-3zf(+/-) vectors to facilitate sequencing. The nucleotide sequence of this PCR product showed about 74% and 70% identity with the same regions of the oat and tomato ADC cDNA sequences, respectively. The predicted .amino acid sequence exhibited 45% and 62% identity with oat and tomato ADC polypeptide fragments, respectively. The sequence similarities of 34%, 47%n and 38% were previously reported in oat and E. coli, tomato ;md oat, and tomato and E. coli ADC amino acids, respectively. Therefore, similarities and identities between rice and oat or tomato are remarkably higher than those others of the previous reports. In the highly conserved regions in both the amino acid sequence and spacing regions among the sequences of these three, rice ADC open reading frame also has the exactly same regions with the striking similarity. RNA blot analysis showed that ADC is expressed as a transcript of approximately 2.5 kbp in the rice seedling leaf tissues.
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