Expression of full-length human pro-urokinase in mammary glands of transgenic mice |
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Authors: | Yaroslav Gursky Robert Bibilashvili Mikchail Minashkin Alex Krasnov Alex Deikin Tatyana Ermolkevich Andrey Popov Lilia Verbovaya Nicolai Rutkevich Alexsander Shevelev Sofia Georgieva Sergey V Razin Igor Goldman Elena Sadchikova |
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Institution: | (1) Institute of Gene Biology Russian Academy of Sciences, Vavilova 34/5, 119334 Moscow, Russia;(2) Institute of Experimental Cardiology, Russian Cardiology Research and Development Centre, 121552 Moscow, Russia |
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Abstract: | Human pro-urokinase expressed in the mammary glands of transgenic animals is quickly activated and converted to urokinase
by proteases that are present in the milk. Thus, it is nearly impossible to isolate full-sized pro-urokinase from the milk
of transgenic animals. To solve this problem, we constructed transgenic mice that express human pro-urokinase and modified
ecotin, which is a potent serine protease inhibitor from E. coli, in their mammary glands. The gene encoding ecotin was modified so as to enhance its specificity for the human urokinase-type
plasminogen activator. Co-expression of modified ecotin and human pro-urokinase in the mammary glands allows for purification
of full-length human pro-urokinase from these transgenic mice. The results described here suggest a general way of preventing
the activation of zymogens that are expressed in the mammary glands of transgenic animals by co-expression of a zymogen along
with a protease inhibitor. |
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Keywords: | Pro-urokinase Ecotin Transgenic animals Milk Inhibitor Protease Thrombolytic agents |
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