Cloning and characterization of a cDNA encoding the chloroplastic copper/zinc-superoxide dismutase from pea.
A cDNA encoding the chloroplastic copper/zinc-superoxide dismutase of pea (Pisum sativum L.) was isolated from a cDNA library constructed in lambda gt11 from leaf mRNA. Nucleotide sequence analysis of the 875-base-pair clone revealed that it contained the complete coding sequence of the mature superoxide dismutase isozyme subunit, along with sequence information for a 48-amino acid N-terminal transit peptide. The deduced amino acid sequence of the mature subunit proved to be 64-87% homologous with amino acid sequences of copper/zinc-superoxide dismutases from other plant species. In vitro transcription, followed by cell-free translation, of the cDNA resulted in the formation of a 23.5-kDa precursor polypeptide, which, upon incubation with isolated pea chloroplasts, was imported and processed to its mature subunit molecular mass of 17.4 kDa.
Scioli
J R
JR
Department of Biochemistry and Microbiology, Cook College, Rutgers University, New Brunswick, NJ 08903.
Zilinskas
B A
BA
D000595:Amino Acid Sequence; D001483:Base Sequence; D001937:Brassica; D002736:Chloroplasts / Q000201:enzymology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004262:DNA Restriction Enzymes; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae; D007527:Isoenzymes / Q000235:genetics*; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology / Q000235:genetics*; D010946:Plants, Medicinal; D014176:Protein Biosynthesis; D013482:Superoxide Dismutase / Q000235:genetics*; D014158:Transcription, Genetic; D014675:Vegetables; D003313:Zea mays
D007527:Isoenzymes; D004247:DNA; D013482:Superoxide Dismutase; D004262:DNA Restriction Enzymes