Meeting Abstract
The clottable protein (CP) is molecules involve in the blood coagulation systems and an essential immune mechanism of crustaceans. Interestingly, CPs in decapod crustaceans exhibit high sequence similarity to vitellogenins (Vgs) in other invertebrates. Previously, single copy of CP (Liv-CP1) was isolated in L. vannamei. In the present study, we identified the additional full-length cDNA encoding the putative CP (Liv-CP2) from the same shrimp by the combination of next-generation sequencing (NGS) and typical PCR-based cloning strategy. Obtained Liv-CP2 exhibited the highest similarity to CP from Marsupenaeus japonicus (33%) and L. vannamei (33%). The newly obtained Liv-CP2 contained the special regions involved in the clotting reactions: Lys-rich and Gln-rich regions, Ser-Lys-Thr repeats, conserved N-glycosylation sites suggesting it may exhibit similar function to Liv-CP1. Phylogenetic analysis showed that the CPs from decapod crustaceans were clustered together with Vgs from insects and vertebrates. Transcriptional analysis of three genes (Liv-CP1, Liv-CP2, and Liv-Vg) was also determined according to the molting cycle and maturation stages by qualitatively and quantitatively. In order to estimate the functions of the newly identified Liv-CP2, RNAi experiments with sequence-specific dsRNA were performed and its physiological effects were also determined.
