1987•Proceedings of the National Academy of SciencesOpen access

Cloning of cDNA for maize superoxide dismutase 2 (SOD2).

Ronald E. Cannon, Joseph Robert White, John G. Scandalios

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Abstract

A cDNA clone encoding maize cytosolic superoxide dismutase 2 (SOD2) was isolated from a cDNA library constructed in pUC9 plasmids from size-selected poly(A)+ RNA. The library was screened with mixed synthetic oligode-oxynucleotide probes. The sequence of the probes was derived from the amino acid sequence from a region of the protein near the NH2 terminus. One positive clone contained an insertion of a 612-base-pair fragment. The amino acid sequence, deduced from the nucleotide sequence of the cDNA, revealed that the clone contained the coding region for all but the first of the 151 amino acids of the SOD2 protein. Additional 5' and 3' flanking sequences, absent on the pUC9 Sod2 clone, were obtained from a lambda gt11 clone isolated from a maize leaf library probed with the pUC9 Sod2 insert. Hybrid-selection translation assays also demonstrate that the cDNA clone contains Sod2 sequences.

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What this paper is about

A cDNA clone encoding maize cytosolic superoxide dismutase 2 (SOD2) was isolated from a cDNA library constructed in pUC9 plasmids from size-selected poly(A)+ RNA. The library was screened with mixed synthetic oligode-oxynucleotide probes. The sequence of the probes was derived from the amino acid sequence from a region of the protein near the NH2 terminus. One positive clone contained an insertion of a 612-base-pair fragment. The amino acid sequence, deduced from the nucleotide sequence of the cDNA, revealed that the clone contained the coding region for all but the first of the 151 amino acids of the SOD2 protein. Additional 5' and 3' flanking sequences, absent on the pUC9 Sod2 clone, were obtained from a lambda gt11 clone isolated from a maize leaf library probed with the pUC9 Sod2 insert. Hybrid-selection translation assays also demonstrate that the cDNA clone contains Sod2 sequences.

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Available abstract

A cDNA clone encoding maize cytosolic superoxide dismutase 2 (SOD2) was isolated from a cDNA library constructed in pUC9 plasmids from size-selected poly(A)+ RNA. The library was screened with mixed synthetic oligode-oxynucleotide probes. The sequence of the probes was derived from the amino acid sequence from a region of the protein near the NH2 terminus. One positive clone contained an insertion of a 612-base-pair fragment. The amino acid sequence, deduced from the nucleotide sequence of the cDNA, revealed that the clone contained the coding region for all but the first of the 151 amino acids of the SOD2 protein. Additional 5' and 3' flanking sequences, absent on the pUC9 Sod2 clone, were obtained from a lambda gt11 clone isolated from a maize leaf library probed with the pUC9 Sod2 insert. Hybrid-selection translation assays also demonstrate that the cDNA clone contains Sod2 sequences.

Key concepts: Complementary DNA, cDNA library, SOD2, Biology, clone (Java method), Molecular biology, Peptide sequence, Nucleic acid sequence

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