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The rusticyanin gene from the acidophilic chemolithotroph Thiobacillus ferrooxidans has been cloned and sequenced. A central portion of the gene was identified by PCR reactions utilising primers optimised for codon bias followed by nested PCR with degenerate primers. The 5 and 3 ends of the rusticyanin gene were then cloned using degenerate primers to each end and anchor sequences to the known...
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