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To facilitate Gibson assembly into the pET-28a(+) vector, short sequences were added at the 5 end (CTCTAGAAATAATTTTGTTTAACTTTAA-GAAGGAGATATACC) and 3 end (GATCCGGCTGCTAACAAAGCCCGAAAG) of each gene

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The initial interpretation of these data was that they represented support for the presence of an NADPH-oxidase in spermatozoa that could be responsible for the high rates of NAD(P)H-induced, lucigenin-dependent chemiluminescence recorded in defective sperm populations [37]