Structure and characterization of a cDNA clone for phenylalanine ammonia-lyase from cut-injured roots of sweet potato
- National Institute of Agro-Environmental Sciences, Ibaraki (Japan) Univ. of Tokyo (Japan)
A cDNA clone for phenylalanine ammonia-lyase (PAL) induced in wounded sweet potato (Ipomoea batatas Lam.) root was obtained by immunoscreening a cDNA library. The protein produced in Escherichia coli cells containing the plasmid pPAL02 was indistinguishable from sweet potato PAL as judged by Ouchterlony double diffusion assays. The M{sub r} of its subunit was 77,000. The cells converted ({sup 14}C)-L-phenylalanine into ({sup 14}C)-t-cinnamic acid and PAL activity was detected in the homogenate of the cells. The activity was dependent on the presence of the pPAL02 plasmid DNA. The nucleotide sequence of the cDNA contained a 2,121-base pair (bp) open-reading frame capable of coding for a polypeptide with 707 amino acids (M{sub r} 77,137), a 22-bp 5{prime}-noncoding region and a 207-bp 3{prime}-noncoding region. The results suggest that the insert DNA fully encoded the amino acid sequence for sweet potato PAL that is induced by wounding. Comparison of the deduced amino acid sequence with that of a PAL cDNA fragment from Phaseolus vulgaris revealed 78.9% homology. The sequence from amino acid residues 258 to 494 was highly conserved, showing 90.7% homology.
- OSTI ID:
- 6874152
- Journal Information:
- Plant Physiology; (USA), Vol. 90:4; ISSN 0032-0889
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
LYASES
DNA-CLONING
POTATOES
INJURIES
AMINO ACID SEQUENCE
CARBON 14 COMPOUNDS
CHEMICAL COMPOSITION
CINNAMIC ACID
COMPARATIVE EVALUATIONS
ENZYME ACTIVITY
MOLECULAR STRUCTURE
PHASEOLUS
PHENYLALANINE
TRACER TECHNIQUES
AMINO ACIDS
CARBOXYLIC ACIDS
CLONING
DNA HYBRIDIZATION
ENZYMES
FOOD
HYBRIDIZATION
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LEGUMINOSAE
MAGNOLIOPHYTA
MAGNOLIOPSIDA
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
PLANTS
TUBERS
VEGETABLES
550201* - Biochemistry- Tracer Techniques