Studies On The Differential Proteome Of Schistosomula Of Schistosoma Japonicum And The Cloing,Expressing Of Schistosomula Differential Expressed Gene SjEnol

Schistosomiasis japonicum is an important parasitic zoonosis in China. There are significant differences in protein expression and biophysicochemical characteristics between schistosomulum and adult male as well as adult female worm. Schistosomulum is the early development stage of schistosome in the final host, some of its differentially expressed proteins may be important in the growth of Schistosoma japonicum. In this paper, the differentially expressed proteins of schistosomulum were identified by proteiomic techniques, the gene encodes the differentially expressed protein SjEnol was cloned and expressed in E coli, the studies provide foundation for the development of effective molecular vaccine, diagnositic antigen and new drug for the control of S. japonicum.Two-dimansional gel electrophoresis was employed to separate soluble proteins from schistoso -mulum(16 days) as well as the male and female worm (40 days) of Schistosoma japonicum, the maps were analyzed by software Image-master. There were 2000 + 69, 1100±75 and 1310+18 spots that were detected in schistosomulum and male as well as female worm respectively, and the molecular weight of most proteins being around 10-80 KD, and PI around 3~8. 26 + 2 proteins were found differentially expressing in schistosomulum.Ten schistosomulum differentially expressed proteins were picked to be further analyzed by mass spectrometry. The purified proteins were identified by software Date MASCOT, Profound and Scope after MOLD-TOFF. The result showed that the predicted functions of these differentially expressed proteins are related to transcription, translation and other important biological fuction of Schistosoma japonicum.A differentially expressed protein of Schistosomulum Enolase (SjEnol) was chosen for further studies. The partial amino acid sequence of this protein was obtained by ESI. The gene containing the ORF sequence was obtained by RACE and PCR, which has 1305 bp that can be translated into 434 amino acids. Recombinant plasmid pET28 a (+) -SjEnol was constructed and expressed in E.coli. The...