Cloing、Expression And Evaluation Of Caspase9of Schistosomiasis Japonicum

Schistosomiasis is a zoonotic disease that is widely spread in the world.Our country is mainly Schistosoma japonicum. For many years,Schistosomiasis prevention and control work has made remarkable achievements,mainly using Snail control, environmental improvement, health education, and praziquantel,but in our country, Many of our towns is still endemic areas of schistosomiasis.Relying only on the combination of these measures and a single drug therapy is difficμLt to eradicate schistosomiasis,Therefore, strengthening the schistosome vaccine development and manufacture new drugs is undoubtedly an important measure for prevention of schistosomiasis.Caspase9is an importent protein in Apoptosis pathway,which Can promote the body’s tissues and organs of apoptosis.In the research,sjcaspase was cloned,expression in Ecoli, and the immuno protective efficiency was evalued. For the depth of sjCaspase biological functions, evaluating it as a vaccine candidate molecμLes and drug targets provide the basis of applications.Before this reserch,Our laboratory has done Do different sources of parasite host Differential gene expression analysis,which found different gene expression among Susceptible host mice, non-susceptible host rats and Microtusfortis which is not the only incidence of schistosomiasis infection.Caspase9was a number of them.Basing on the study, Reference SmCaspase9gene amplification method,acquiring complated Sjcaspase9cDNA sequence. Bioinformatics analysis, the Open Reading Frame for Sjcaspase9was1140bp, encoding380amino acids with a molec μ Lar weight of44kD。The two recombinant proteins had antigenicity and immunogenicity revealed by western blotting. Using Purified SjCaspase9-pET-28a recombinant protein immunized BALB/c mice, the resμLts of immunohistochemistry compared with the control group, the recombinant protein SjCaspase9-pET-28a were induced in mice worm reduction rate of26.44%and32.98%liver egg reduction rate, significant difference (p<0.05) Serum specific IgG antibody level changing was detected by ELISA. The resμLts showed that the recombinant protein can induce specific IgG antibodies in mice rapidly produce and vaccine candidates and new potential drug targets as well as some in-depth study of the value. maintain a high level. Indicates that the development of recombinant protein anti-schistosomiasis.Extracted 7d,14d,21d,28d,35d,42d and42d each period of the parasite protein male and female, to a-tublin as an internal, through various periods of parasites detected by Western blotting in Caspase9protein expression. To housekeeping gene a-tublin as an internal, fluorescence quantitative PCR analysis sjCaspase9in rats, mice, worms and mice, Microtus fortis collection period to collect the other larvae in the different transcription conditions. Prove that the gene in the schistosome cercariae infect the host after7d,14d,21d,28d,35d,42d are expressed, including expression of7d the highest expression of male than female. In the East voles, rats, mice also were expressed, the highest expression of oriental voles, rats, followed by mice was the lowest. Caspase9high expression may be part of organ development and parasite-related organ degeneration. The study also built ping pCDNA3.1(+)-Sjcaspase9plasmid, and transfected into cells that can express. In this study, in-depth study of the biological function of genes SjCaspase9laid the foundation for the screening of new molecμLes and schistosomiasis vaccine candidate drug target provides a new idea.