The Function Preliminary Analysis Putative Calcium Binding Protein(TGME49＿229480)of Toxoplasma Gondii

Toxoplasma gondii is an obligate intracellular protozoon that belongs to the Apicomplexa group which can infect nucleated cell.People infected because food contaminated / infected by T.gondii oocysts or cysts.In the intermediate host,tachyzoites and bradyzoites reproduce mainly by endodyogeny.The disease can cause miscarriage and stillbirth in pregnant and pregnant animals.Mortality of more than 60% can be observed in pigs infected with T.gondii.In recent years,with the increasing infection rate of Toxoplasma gondii,developing an effective anti-Toxoplasma drug has become an urgent problem to be solved.Calcium ions as a second messenger is involved in the regulation of multiple pathways.The CaBPs family includes Calmodulin family,Calcineurin B-like and calcium-dependent protein kinase.The Calmodulin family binding to Myo H relation to the process of invasion host cell and physiological activities of Toxoplasma gondii.In this subject,genome of Toxoplasma gondii was predicted and analyzed by bioinformatics,which founded pCaBP have highly conserved domain in different virulence strains and contains multiple calcium-binding domains.It is similar to structure of Ca Ms.We selected pCaBP as target protein,and analyzed whether its bind to the Myo H protein and how does the protein play a role in the invasion of host cells.So,this topic has carried out depth research its function.Two prokaryotic expression vectors were successfully constructed: p GEX-4T-1-pCBP and p ET-28a-CaBP.The pCaBP was localized in cytoplasm of RH strain and ME49 strain by indirect immunization fluorescence.These proteins have different transcription and expression levels,and the expression level of RH strain was low.The molecular interaction experiments revealed that the pCaBP protein binds to Myosin H protein.The above experiments laid the foundation for the further study of the function of pCaBP protein.