Langner K D, Bird R E, McCandliss R, Huber B, Amann E, Zettlmeissl G, Küpper H A
Research Laboratories of Behringwerke AG, Marburg/Lahn, W. Germany.
Behring Inst Mitt. 1988 Apr(82):16-25.
The recent cloning and sequence analysis of human factor VIII:C (antihaemophilic factor) revealed a domain structure for the protein which can be presented as A1-A2-B-A3-C1-C2. In this report we describe the construction of two altered factor VIII:C cDNAs coding for molecules in which a part (amino acids 816 to 1598) or all of the B domain (amino acids 741 to 1689) was removed. In the latter mutant a new thrombin cleavage site has been created, which does not exist in wild-type factor VIII:C. The mutated cDNAs were cloned into eucaryotic expression vectors based on regulatory sequences of the virus SV40 and transfected into two different mammalian cell lines. Both truncated recombinant factor VIII:C molecules were secreted into the culture medium, showed full biological activity and could be activated by thrombin.
近期对人凝血因子VIII:C（抗血友病因子）的克隆及序列分析揭示了该蛋白质的结构域组成，可表示为A1-A2-B-A3-C1-C2。在本报告中，我们描述了两种经过改造的凝血因子VIII:C cDNA的构建，它们编码的分子分别缺失了部分（氨基酸816至1598）或全部B结构域（氨基酸741至1689）。在后一种突变体中，产生了一个野生型凝血因子VIII:C中不存在的新的凝血酶切割位点。将突变的cDNA克隆到基于病毒SV40调控序列的真核表达载体中，并转染到两种不同的哺乳动物细胞系中。两种截短的重组凝血因子VIII:C分子均分泌到培养基中，具有完全的生物学活性，且能被凝血酶激活。
