Efficient Expression Of Genetic Engineering Lumbrokinase And Examination Of Bio-activation

The purpose of the work is to establish an efficiency express Goat Mammary gland bioreactor of lumbrokinase by genetic engineering way, and lumbrokinase can be expressed in milk, finally to check its function. To drink milk and treat thrombus with the purpose of health. Now, three works have been done: lumbrokinase's espression in mammal cells; The acute toxic effects of genetic engineering lumbrokinase; to check antithrombus function of genetic engineering lumbrokinase.To constract pbLK and pb-LK-tpa specialized vector expresses in mammary gland. Then the two expression vectors were injected in goat mammary gland. Fibin agarose plate assay indicated both of the two plasmides expressed in milk. The expression quantity of pIbCP-LK-tpa was higher than pIbCP-LK. It was the first time that lumbrokinase expressed in goat mammary gland and it provided a dependable guarantee for the establishment of transgenic goat and the expression of lumbrokinase in mammary gland.The acute toxic effects of genetic engineering lumbrokinase were evaluated .The routine acute toxic test on mice was performed by perfusing the mice with genetic engineering lumbrokinase. Results: The maximum tolerance dosage of mice in genetic engineering lumbrokinase through mouth was 4.002W IU/kg BW; no microscopic changes were found in the heart, liver, spleen, lung and kidney. It is proved that genetic engineering lumbrokinase had no acute atoxicity.Effect of genetic engineering lumbrokinase on Thrombus Formation in the Arteries in Rats. Methods to observe the thrombosis and fibrinolysis effect of OT in the arteries through electric stimulation of common carotid artery. In vivo and in vitro thrombosis, and in vivo fibrinolysis tests were used to evaluate the effects of the drug on the fibrinolysis system. To observe anti- thrombus function were examined in the normal saline group and high-dose, middle-dose, low-dose groups of genetic engineering lumbrokinase. Results High-dose, middle-dose, and low-dose groups of genetic engineering lumbrokinase could prolong OT and decrease Thrombus Formation. it reduced thrombus weight and length significantly; it also showed a strong enhancement on fibrinolysis. Conclusion genetic engineering lumbrokinase could significantly inhibit the formation of thrombus in the arteries, and also increase fibrinolysis.