Extraction And Purification Of Expressed Products HTNF-α From Anabaena 7120

Cyanobacteria are autotrophic and oxygenic photosynthesis microbe. It can be alive under the condition of light, carbon dioxide, inorganic salt and optimal temperature, that's to say, its culture conditions are simple and easy to culture in mass. Preparing hTNF-αhealth product taken orally with the host of cyanobacteria, can avoid the drawbacks of intro toxin and inclusion bodies formation, and solve the problems met in vein injection.Human tumor necrosis factor-α(hTNF-α) is a selective and direct scavenger against tumor tissues and tumor cells. hTNF-αwould be rank among the most promising anticarcinogens in the future.Transgenic hTNF-αAnabaen IB02 were cultured in 5L bottle and disrupted and treated with a series of methods and got the optimal purified conditions. The results showed as follows:1. Transgenic hTNF-αAnabaen sp.PCC IB02 were cultured in 5L bottle which was filled with 4L liquid under 90umol/m2.s-1 light and ascertained the optimal collected time of algas at 24th day.2. Screened out the optimal extracted buffer from a serials of buffers and cell disrupture methods. The results showed that: the optimal dissolving conditions of hTNF-α: 0.05mol/L, pH7.5 Tris-HCl, the concentration of NaCl: 0.2 mol/L, of EDTA: 0.01mol/L; The best conditions of supersonic: supersonic power 800W, interval time 1s, run time 0.5s, the ratio of the mass of alga to volume of disrupting buffer 1:15, supersonic time 8min, disrupting one time. And the best conditions of freezing-thawing: volume of container: <1.5mL, firstly, freezed the whole night at -20℃, centrifuged and got the supernatant after thawing, then added the same volume of buffer and went on freezing for two hours, got the supernatant, at last mixed supernatant.