The cDNA Sequence of Two Hemocyanin Subunits from Red Swamp Crayfish Procambarus clarkii and their Responses to White Spot Syndrome Virus Infection.

Hemocyanin, the respiratory protein of crustaceans, participates in the innate immune defense in these organisms. We cloned two hemocyanin subunit genes (PcHc1 and PcHc2), by using a degenerate primer PCR and rapid amplification of cDNA ends (RACE) approach, from the hepatopancreas of red swamp crayfish Procambarus clarkii. The transcripts of these two subunits were only detected in the hepatopancreas by reverse transcriptase quantitative PCR (RT-qPCR) analysis. The neighbor-joining and maximum parsimony phylogenetic analyses indicated that PcHc2 associated with a clade belong to the α-type hemocyanins and PcHc1 associated with another clade belonging to the β-type hemocyanins. The data obtained from the RT-qPCR indicated that the mRNA expression levels of these subunit genes followed almost the same regulation pattern in the crayfish challenged with white spot syndrome virus (WSSV). The fluctuation of mRNA expression levels of these two subunits after the WSSV challenge indicated that both of them may participate in the antiviral immune response of crayfish. Received April 12, 2015; accepted November 22, 2015.