High Density Fermentation For Genetically Engineered Pichia Pastoris And Cloning Of Two Chicken Cytokines

Interferons(IFNs), a family of cytokines, plays an important role in many of biological activities, such as interfering with the replication of various viruses, decreasing cell proliferation and modifying immunological processes. IFNα/β is the first system of body to defend virus attacking. Pig production is important for food industry, but many swine immuno-suppresive diseases are getting more and more popular nowdays. Therefore, recombinant Porcine- IFN α is desirable as immuno-activation and immunotherapy agent.In this study, the optimum culture conditions of genetically engineered Pichia pastoris in shake-flask cultivation and in fed-batch fermentation were reported respectively in this paper. Experimental results revealed that the culture period induced with methanol was 96 h. The optimum methanol concentration was 8g/L, pH range was 6.4-9.0 and the seed inoculum amount was 2:1 in shake-flask cul:ure. Altnough the seed inoculum amount increased, the target protein concentration was decreased. The 10% inoculum and 5-6 OD₆₀₀ of seed were beneficial to the high cell density cultivation . With glycerol feeding strategy based on DO level, the cell dry weight of broth reached 118. 7g/L and the IFN concentration was 1. 034g/L by methanol inducement for 48 hours at thefed-batch fermentation phase.Part Two C1ONING OF TWO CHICKEN CYTOKINESIt has been proved that recombinant animal cytokines can defend the pestilence and reduce the graziery losing which caused by diseases effectively. Therefor it has great ecnomic value. The research of animal cytokines become popular on the preclinical and clinical study, such as their mocecular structure,...