Abstract
In order to clone a Cu/Zn-SOD gene from Aspergillus japonicus, the highly conserved regions of the SOD sequence of maize and other species were compared and two oligonucleotides were synthesized. The Aspergillus japonicus cDNA was then used as the template to get a 0.3 kb fragment by PCR technique. The 0.3kb DNA fragment has high homology compared with maize SOD-4 cDNA. Then we used the 0.3kb fragment as probe to screen Aspergillus japonicus genomic libraries. A 0.4 kb DNA frgment from purified positive plaque was subcloned into pGEM-7zf(+) vector and sequenced. Nucleotide sequence analysis of this clone revealed that it comprised an open reading frame coding for 120 amino acid residues. The encoded polypeptide is highly homologous with other plant SODs. The residues required for coordinating copper and zinc are also conserved as they are among all reported Cu/Zn-SOD sequences.
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