Clone Expression And Analysis Of The Schistosoma Japonicum Sjwnt11, Sjwnt5and Sjwnt1

Praziquantel is the only chemotherapy drug to treat the disease. Satisfactory therapeuticeffect, but the long-term single, repeated drug use, there is a risk of drug resistance[1,2]. Fullyunderstand the regulation of growth and development of the schistosome mechanism is the basisof the research and development of new drugs. For the study of Schistosoma japonicum Wntsignal transduction protein gene developmental regulation of the parasite,on the various stagesand sex of Wnt mRNA expression were analyzed.23days parasite cDNA as a template, whichamplified protein and nucleotide sequences of SjWnt11, SjWnt5SjWnt1, and constructedrecombinant expression plasmid of PET28a-SjWnt11, PET28a-SjWnt5and PET28a-SjWnt1,and conduct a protein expression and purification.The purified wnt gene recombinant proteinimmunized BALB/c mice, and specific polyclonal antibody was prepared for the follow-upstudy. Using Western blot analized expression levels of SjWnt5proteins in differentdevelopmental stages of the parasite. Use GenomeWalking technical analysis of the geneticstructure of Schistosoma japonicum SjWnt11.23days genital initially built started and SjWnt5mRNA expression decreased in females, while in males constantly increased.SjWnt11mRNA expression at13days the increase, which presumably SjWnt11may adjustprocess occurs of parasites organ. It inferred SjWnt5triggered Wnt signaling had a moderatingeffect for the early development of organ differentiation parasites. The completion of the femalereproductive organs, and the further development of sexually mature males are likely to haveregulatory role. The mRNA of SjWnt1were the highest expression in7days, and eggs periodwere the second expression.13days later,Period expression remained at the lowest level. Itspeculated that the protein in the egg parasites early embryo development and organdifferentiation play a regulatory role in the process. SjWnt4mRNA level was the highest at7days schistosomula of expression, and in other developmental stages of expression wasrelatively low. The result suggested that SjWnt4and embryo development, body early organdifferentiation. SjWnt2was also expressed in different at developmental stages of Schistosomajaponicum differences. The highest expression level was in the egg period, and7daysschistosomula cell differentiation expression was down-regulated. This was probably related toembryo formation. By comparing the binding receptor frizzled mRNA in different period ofgender expression, SjWnt11and frizzled7, SjWnt1, frizzled5and frizzled8, SjWnt4andfrizzled9expression tendency was consistent.It speculated that SjWnt11and frizzled7may interact, SjWnt4and frizzled9may interact, SjWnt1, frizzled5and frizzled8may interact. Onthe25day sex infection of male and female worms and multiple infection of male and femaleworms, There was a big difference between the level of Wnt gene expression of mRNA. It canbe speculated that The Wnt signal may play a role in the regulation of the female reproductiveorgan built, further development and maturation of male, may play a role in the regulation ofWnt signal. The successful construction of SjWnt11, prokaryotic expression of SjWnt5andSjWnt1gene plasmid was expressed successfully in e.coli, and polyclonal antibody preparationcan be single and the larvae of natural protein with reaction. This proves that polyclonalantibody had good specificity. By Western blot method, comparison of Schistosoma japonicumSjWnt5in protein level in different developmental stages of the differences, the trend of theexpression of SjWnt5protein and SjWnt5mRNA was consistent. Combined with thedevelopment characteristics of Schistosoma japonicum, which furtherly confirmed that SjWnt5gene plays an important role in the process of Schistosoma japonicum organ differentiation.Andgene structure of SjWnt11were obtained, with5introns.Schistosoma japonicum SjWnt11, SjWnt1and SjWnt5had been analysed in Various stagesand gender mRNA. Comprehensive analyed that Wnt pathway may be involved in thedevelopment of Schistosoma japonicum multiple conditions, such as cell differentiation, organformation, male and female reproductive organs built and maturation and embryo development.Studying for the further use of RNA interference technique, Wnt gene and provide basic data toregulate the development of Schistosoma japonicum.