Cloning and Anti-bacterial Functional Analysis of EsPrx4 in Eriocheir sinensis
LIANG Meng1,2*, WANG Mei-Yao1,2*, LI Jian-Lin1,2, YAN Feng-Yuan3, ZHOU Jun4, TANG Yong-Kai1,2,3**
1 Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214128, China; 2 Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China; 3 College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China; 4 Freshwater Fisheries Research Institute of Jiangsu Province, Nanjing 210017, China
Abstract：In recent years, deterioration of water environment, elevation of stocking density, occurrence of disease and abiotic stress have seriously affected growth and development of Eriocheir sinensis. Peroxiredoxin (Prx) plays an important role in oxidative stress response. Prx4 is an important member of Prx family. To discuss the role of Prx4 in the anti-bacterial response of E. sinensis, EsPrx4 (GenBank No. ON685195) was cloned from hepatopancreas with homologous cloning method. Sequence analysis showed that the coding region of EsPrx4 was 738 bp, encoding 245 amino acids. The physical and chemical propertiese analysis showed that the molecular weight of EsPrx4 protein was 27.31 kD, the theoretical isoelectric point was 5.55, and the total average hydrophilicity was -0.181, indicating that EsPrx4 protein was hydrophilic. Phylogenetic analysis showed that the EsPrx4 has high homology to that of Scylla paramamosain, Penaeus monodon and Litopenaeus vannamei, which indicated high conservation in Prx4 of shrimp and crab. Results of qPCR indicated that EsPrx4 was widely expressed in E. sinensis, it had the highest expression in hepatopancreas and the lowest expression in intestine. Under the stimulation of Listonella anguillarum, the expression of EsPrx4 reached the highest level at 24 h and recovered to the normal level at 72 h, indicating that EsPrx4 plays an important role in the immune response of E. sinensis. This study provides theoretical support for further research on innate immunity and Prx gene family of E. sinensis.
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