Cloning And Expression Analysis Of Immune-related Genes In Exopalaemon Carinicauda

The Exopalaemon carinicauda is one of the most important commercial shrimpspecies in Yellow Sea and Bohai Sea, China. However, with development of intensiveculture, the culturing environment grow polluted and various diseases caused bybacteria and viruses have blossomed within booming E. carinicauda cultures, that makegreat losses to commercial shrimp aquaculture. To understanding the innate immuneabilities and immune defense mechanisms of shrimp, we will be beneficial to thedevelopment of health management and disease control in shrimp aquaculture.Base on the hemocytes cDNA library of E. carinicauda in our lab, thealpha2-macrogloblin cDNA of Exopalaemon carinicauda (named Ec2M) was clonedby rapid amplification of cDNA ends (RACE). The Ec2M cDNA was4823bp inlength, which contains an open reading frame (ORF) of4413bp, a64bp5’untranslatedregion, and a346bp3’untranslated region. The open reading frame encodes anN-terminal signal sequence of23residues, and a mature protein of1470amino acids,with the molecular mass of163.0kDa and an estimated pI of5.03. Homology analysisrevealed that the amino acid sequence of Ec2M was highly conserved with itshomologs in other crustaceans. The similarities of Ec2M with the Ec2M ofMacrobrachium rosenbergii were80%. The expression levels of Ec2M in differenttissues were analyzed by quantitative real-time PCR. Expression of Ec2M wasdetected in all tested tissues of E. carinicauda, including hemocytes, gill,hepatopancreas, muscle, ovary, intestine, stomach and eyestalk, with the highestexpression level in hemocytes. After challenged with Vibrio anguillarum and white spotsyndrome virus, the expression of Ec2M were up-regulated in the hemocytes andhepatopancreas of E. carinicauda. Our results implied that Ec2M might play animportant role in the prawn immune response.Base on the hemocytes cDNA library of E. carinicauda in our lab, the14-3-3cDNA of Exopalaemon carinicauda (named Ec14-3-3) was cloned by rapidamplification of cDNA ends (RACE). The Ec14-3-3cDNA was2905bp in length,which contains an open reading frame (ORF) of744bp, a127bp5’untranslated region,and a2033bp3’untranslated region. The open reading frame encodes a mature proteinof247amino acids, with the molecular mass of27.95kDa and an estimated pI of4.65.Homology analysis revealed that the amino acid sequence of Ec14-3-3was highlyconserved with its homologs in other crustaceans. The similarities of Ec14-3-3with the Penaeus monodon were98%. The expression levels of Ec14-3-3in different tissueswere analyzed by quantitative real-time PCR. Expression of Ec14-3-3was detected inall tested tissues of E. carinicauda, including hemocytes, gill, hepatopancreas, muscle,ovary, intestine, stomach and eyestalk, with the highest expression level in hemocytes.After challenged with Vibrio anguillarum and white spot syndrome virus, the expressionof Ec14-3-3were up-regulated in the hemocytes and hepatopancreas of E. carinicauda.Our results implied that Ec14-3-3might play an important role in the prawn immuneresponse.Base on the hemocytes cDNA library of E. carinicauda in our lab, the C-typelectin cDNA of Exopalaemon carinicauda (named EcCTL) was cloned by rapidamplification of cDNA ends (RACE). The EcCTL cDNA was1285bp in length, whichcontains an open reading frame (ORF) of1041bp, a62bp5’untranslated region, and a182bp3’untranslated region. The open reading frame encodes a mature protein of346amino acids, with the molecular mass of38.56kDa and an estimated pI of4.29.Homology analysis revealed that the amino acid sequence of EcCTL was highlyconserved with its homologs in other crustaceans. The similarities of EcCTL with thePalaemon modestus were91%. The expression levels of EcCTL in different tissueswere analyzed by quantitative real-time PCR. Expression of EcCTL was detected in alltested tissues of E. carinicauda, including hemocytes, gill, hepatopancreas, muscle,ovary, intestine, stomach and eyestalk, with the highest expression level inhepatopancreas. After challenged with Vibrio anguillarum and white spot syndromevirus, the expression of EcCTL were up-regulated in the hemocytes and hepatopancreasof E. carinicauda. Our results implied that EcCTL might play an important role in theprawn immune response.