HEMORRHAGIC AND NECROTIC TOXINS IN SNAKE VENOMS

蛇毒中的出血性和坏死性毒素

基本信息

批准号：
2291672
负责人：
ANTHONY T TU
金额：
$ 2.45万
依托单位：
COLORADO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-09-30 至 1997-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/2291672
关键词：
Baculoviridae animal genetic material tag anticoagulants complementary DNA drug design /synthesis /production fibrinolytic agents gel filtration chromatography genetic library immunoaffinity chromatography messenger RNA protein sequence recombinant proteins snakes thrombin tissue /cell culture toxin transfection /expression vector

项目摘要

Snake venom is a mixture of proteins, toxic and nontoxic components. It has been known that venoms of some Crotalidae and Viperidae have anticoagulation activity. Recent work in our laboratory and also in Dr. Siigur's laboratory isolated fibrinolytic enzymes without toxic action from the venoms of Vipera lebetina (lebetase) and Crotalus Atrox (atroxase). Thrombosis is a leading killer throughout the world, and it is important to find better thrombolytic agents than those currently used such as tpA, urokinase, and streptokinase. Lebetase, atroxase, and other fibrinolytic enzymes from snake venoms are potentially useful thrombolytic agents after the removal of toxic components from snake venoms. It is proposed that base sequence of cDNA encoded for lebetase will be determined. For this purpose mRNA will be first isolated from venom glands of Vipera lebetina. Lebetase gene will be isolated and sequenced. Recombinant lebetase will also be produced by packaging lebetase gene into baculovirus transfer vector. The cloned lebetase will be compared with the enzyme isolated from Vipera lebetina venom. Once cloned lebetase is shown to be identical to the natural one, fibrinogenase activity can be increased by modifying the enzyme in the future. Our proposed work may produce a new potent thrombolytic agent.

蛇毒是蛋白质、有毒和无毒成分的混合物。它众所周知，一些响尾蛇科和蝰蛇科的毒液具有抗凝活性。我们实验室以及博士的最新工作。 Siigur实验室分离出无毒性作用的纤溶酶来自 Vipera lebetina (lebetase) 和 Crotalus Atrox 的毒液（阿托氧化酶）。血栓形成是全世界的头号杀手，找到比目前使用的更好的溶栓剂非常重要例如tpA、尿激酶和链激酶。 Lebetase、阿托酶等蛇毒中的纤溶酶具有潜在用途蛇体内有毒成分去除后的溶栓剂毒液。建议编码 lebetase 的 cDNA 的碱基序列为决定。为此，首先从毒液中分离出 mRNA Vipera lebetina 的腺体。 Lebetase 基因将被分离并测序。重组lebetase也将通过包装lebetase基因来生产转入杆状病毒转移载体。将比较克隆的lebetase 用从 Vipera lebetina 毒液中分离出的酶。一旦克隆了lebetase 被证明与天然纤维蛋白原酶相同，纤维蛋白原酶活性可以将来可以通过修饰酶来增加。我们提出的工作可能生产一种新的强效溶栓剂。