The full-length cDNA of the alternative oxidase (AOX) gene from Porphyra yezoensis Ueda (PyAOX) [currently assigned as Pyropia yezoensis (Ueda) M. S. Hwang et H. G. Choi (http://www.algaebase.org)] an ancient member of the Rhodphyta, was cloned by electronic cloning, rapid amplification of cDNA ends (RACE), and reverse transcription PCR. The nucleotide sequence of PyAOX consists of 1,650 bp, including a 5′ untranslated region (UTR) of 170 bp, a 3′ UTR of 148 bp, and an open reading frame (ORF) of 1,332 bp that can be translated into a 443-amino-acid residue with a molecular mass of 47.33 kDa and a putative isoelectric point (pI) of 9.71. The putative amino acids had 50%–61% identity with AOX genes in Eukaryota and higher plants and had AOX-like characteristics. The expression of PyAOX mRNA in different stages of the life cycle, conchospores, filamentous thalli (conchocelis stage), and leafy thalli, was detected by real-time quantitative PCR (qPCR). The highest level of expression, which was observed in filamentous thalli, was three times higher than that observed in leafy thalli. The next highest level, which was observed in the conchospores, was twice as high as that observed in leafy thalli. We showed that an alternative respiration pathway existed in P. yezoensis with a noninvasive microsensing system. The contribution of the alternative pathway to total respiration in filamentous thalli was greater than that in leafy thalli. This result was consistent with the level of AOX gene expression observed in different stages of the life cycle.