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Diphthamide biosynthesis protein 7 (Dph7) is a vital protein for diphthamide biosynthesis in archaea and eukaryotes. The 1143 bp cDNA sequence of Dph7 was cloned from the gills of Marsupenaeus japonicus using RT-PCR and RACE. Data showed that Dph7 was highly expressed in the gills and digestive gland of M. japonicus. Furthermore, the expression of dph7 was induced by infection with white spot syndrome virus (WSSV). When Dph7 was knocked down, immune genes such as toll, prophenoloxidase (proPO), p53, tumor necrosis factor-α (TNF-α) and signal transducer and activator of transcription (STAT) were significantly down-regulated (P < 0.01) in hemocytes. First, we demonstrated that Dph7 is very important in the progression of WSSV infection and that the time of death for WSSV-infected shrimp was significantly advanced following RNAi targeting of Dph7. We also investigated the effect of Dph7 on apoptosis rate in M. japonicas and found that Dph7-dsRNA treatment caused lower levels of apoptosis in hemocytes, both in the disease-free group and the WSSV group. Knock-down of Dph7 affected the activity of both phenoloxidase (PO) and superoxide dismutase (SOD), and total hemocyte count (THC) after infection with WSSV, indicating that Dph7 plays a regulatory role in the immunological reaction of shrimp in response to WSSV infection. Thus, we conclude that Dph7 may promote the anti-WSSV immune response of shrimp by regulating apoptosis, SOD and PO activity, and can influence the progression of WSSV infection.The 1143 bp cDNA sequence of Diphthamide biosynthesis protein 7 (Dph7) was cloned from Marsupenaeus japonicus.The Dph7 is very important for the progression of WSSV infection.The Dph7 knockdown significantly increased the mortality of WSSV-infected shrimps.The Dph7 might promote the immune response of shrimp by regulating apoptosis, PO activity and SOD activity.