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Vol 47(2013) N 4 p. 544-551; Y. Fang1,2, K. Jiang2, F. Zhang2, M. Sun1,2, J. Hu1,2, L. Ma2* Macrophage migration inhibitory factor in mud crab Scylla paramamosain: molecular cloning, expression profiles in various tissues and under Vibrio challenge 1College of Fisheries and Life Science, Shanghai Ocean University, 999 Huchenghuan Road, Shanghai, 201306, China2East China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, 300 Jungong Road, Shanghai, 200090, China *malingbo@vip.sina.com Received - 2012-06-29; Accepted - 2012-10-29 As one of the first found cytokines, macrophage migration inhibitory factor (MIF) plays important roles in several physiological processes in crabs. In this study, a full-length MIF cDNA (GenBank accession number: JX131610) from mud crab Scylla paramamosain (Sp) was cloned based on a sequence of S. paramamosain cDNA library. The full length of SpMIF was 734 bp consisting of a 363 bp open reading frame encoding the SpMIF, a 120 amino acids peptide chain. The molecular weight of SpMIF was 13.46 kDa with the pI of 6.82. The alignment analysis showed that SpMIF appeared to be closely related to the counterpart from Eriocheir sinensis (68%). Quantitative real-time PCR analyses revealed that SpMIF was highly expressed in hepatopancreas and hemocytes. In addition, the expression level of SpMIF was increased significantly after 6-h challenge by Vibrio parahaemolyticus (4.00×106 CFU/mL), peaked at 8 h, and then declined to the common level in 48 h. These data indicated that SpMIF was cloned successfully, and suggested that it participated in the immune system of mud crabs. macrophage migration inhibitory factor,quantitative real-time PCR,Scylla paramamosain,tissue expression |
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