| Summary: | 碩士 === 國立高雄海洋科技大學 === 海洋生物技術研究所 === 100 === The aim of this study is to identify the shrimp proteins which can bind to the envelop proteins VP28 of white spot syndrome virus (WSSV）. The recombinant VP28 protein was expressed in E. coli BL21 (ED3), purified by affinity column, and used to prepare a probe by coating on magnetic nanoparticles (MNPs). The probe, called VP28@MNP, was then used to screen the corresponding interacting proteins within whole haemolymph (blood) of Litopenaeus vannamei by incubation at 4oC for 30 min. Several regular bands were found in SDS-PAGE and further identified by LC-ESI tandem mass spectrometry after trypsin digestion. These VP28-binding proteins include hemolymph clottable protein, hemocyanin, heat shock cognate 70, heat shock protein 60, arginine kinase, actin and histone H2A.
Within whole haemolymph between uninfected and infected shrimp, no significantly different VP28-binding proteins were found. However, the uninfected shrimp blood aggregate with VP28@MNP, but the infected blood does not. It indicated that some factors of WSSV manipulate the binding between VP28-binding protein and VP28@MNP.
Based on these preliminary data, it is proposed that WSSV enter shrimp cell via the transport of lipid by the binding between VP28 and hemolymph clottable protein. VP28 binds hemocyanin, arginine kinase and histone H2A to weaken the immune system by inhibiting phenoloxidase activities, shutting down the energy supplier and prohibiting DNA repairing, respectively.
|
|---|
