OBJECTIVE To ascertain the expression of human tissue-type plasminogen activator(t-PA)cDNA in hen muscles.test the content and the biological activity of t-PA in yolk.METHODS The expression plasmid of pcDNA3-t-PA.Containing t-PA cDNA,was directly injected into hen muscles.Eggs were collected from the hens after the injection.The expression efficiency was evaluated by Southern blotting.The content by ELISA test and the biological activity of t-PA in yolk by Fibrinolysis test.RESULTS Southern blotting and ELISA results showed that there was t-PA accumulated into yolk on 7th day after the injection of the recombinant plasmid.and the peak occurred on 14th to 21st day.The expression yield was 6.1 mg·L-1.Fibrinolysis of t-PA in vitro indicated that the expression product activated tissue-type plasminogen.On 14th to 21st day after the injection of pcDNA3-t-PA,the activity of t-PA contained in yolk per milliliter corresponded to 37-45.8 μg standard t-PA.CONCLUSION pcDNA3-t-PA could be expressed in vivo,and the expressed product could accumulate into yolk and would be of biological activity.This study shows that the recovery of pharmaceutical proteins from the yolk is an effective method in transgenic chicken bioreactors.
LI Yin-ju;WU Ting-ci;ZHNG Chun-jie;CHENG Xing-cho.
Study on in Vivo Expression of t-PA cDNA Directly Injected into Hen Muscle and Its Biological Activity [J]. Chinese Pharmaceutical Journal, 2008, 43(11): 809-812
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