Expression Of Haemonchus Contortus Hc38Gene In Pichia Pastoris And Biological Characteristion Preliminary Exploration

Hc38gene in the intestinal epithelium microvilli,Australia scholar Hartman(2001) found that this gene present a high expression in the intestinal epithelium microvilli by Northern hybridize,to infer its expression product is a kind of important function of the protein about hemophagia and immune escape, and then provide reliably methods for prevention and cure Haemonchosis. In this study,we have obtained the following results:The porcine HcP gene was amplified from recombinant plasmid Hc38by PCR,then the fusion expression vector pPIC9K-HcP was constructed by inserting HcP fragment, and transformed to GS115by electroporation. Multi-copy recombinant strains were screened by G418. The expression of recombinant fusion protein was induced by methanol, and SDS-PAGE and Western blot were employed for analyzing the product. Fusion protein was purified.The bioactivity of Hc38was measured by hemagglutination assay to cattle,goat,chicken,rabbit,mice; Hcp recombinant protein was incubated with natural host goat’s IgG and hemoglobin,SDS-PAGE was employed for analyzing the digestion condition.The hemagglutination activity of recombinant HcP protein was detected utilizing erythrocytes derived from blood samples of cattle、goat、chicken、rabbit、mice.The results showed that recombinant Hc38protein was blood group specific;meanwhile,recombinant HcP recombinant protein degradation hemoglobin and IgG of the natural host of H.contortus, the goat, proving this gene’s function of the early day’s infer,about hemophagia and immune escape.