Study On Type â…¤ Collagen Of Schistosoma Japonicum

Schistosomiasis is an important global public health problem，but it has been effectively controlledin China. In recent years, with the development of molecular biology and proteomics, the focus ofschistosomiasis researchers gradually shifted to its molecular function and mechanism of life.S. japonicum gynecophoral canal protein (SjGCP) is an important protein molecules in theschistosome life processes, it is a male-specific signaling molecule, which was first discovered in1987by Gupta et al. SjGCP belongs to Fascilin I protein family, it is similar to the periostin protein inmammals. It was proved that periostin protein had an important role in the signaling and mediating ofinsect embryos growth and development. SjGCP have an important role in regulating female sexualmaturity and male and female encircle.Our laboratory using the yeast two-hybrid selected yeast two-hybrid cDNA library of18dS.japonicum, the sequences of kinds of collagen genes interacting with SjGCP was obtained, includingSjColⅤ. In this study, we confirmed the interaction of SjGCP with10S.japonicum proteins, SjColⅤwere selected for further study. The following major progresses were achieved in this work.1. Verified the interaction of ten kinds of Schistosoma japonicum proteins with SjGCP. In thisstudy, the interaction of10positive clones with SjGCP were confirmed. The experimental results shownthat there is interaction between SjColⅤ and SjGCP in the yeast system. To further validate theauthenticity of the interaction, the eukaryotic expression vector of SjColV and SjGCP were constructedand transferred in HEK293T cell. Co-IP technology was adopted to detecte the interaction betweenthem. The results showed that, there is interaction between SjColⅤ and SjGCP.2. To analyze expression of SjColⅤ in E.coli DE3, a prokaryotic expression vector pET28-SjColVwas constructed. rSjColV was expressed in E.coli DE3with induction of IPTG and detected withSDS-PAGE and Western Blot. The results showed that SjColV was efficiently expressed in DE3cells,but the products were unstable. In this study electrospray ionization mass spectrometry（EMS）wasapplied to analyze the prokaryotic expression conditions of SjColV.3. Effects of SjColⅤ and SjGCP silencing on S.japonicum. In this part, RNAi technology wereadopted to silence the expression of SjColⅤ and SjGCP. The results showed that SjColⅤ silencing leadthe spawning and hatching rate of S.japonicum decreased significantly and results in morphologicalchanges of Schistosoma; SjGCP silencing lead to significant changes of the internal structure ofS.japonicum compared with normal worms. It can be speculated that SjColⅤ and SjGCP haveimportant roles in the growth and development of S. japonicum.