Benatti L, Scacheri E, Bishop D H, Sarmientos P
Department of Biotechnology, Farmitalia Carlo Erba, Milan, Italy.
Gene. 1991 May 30;101(2):255-60. doi: 10.1016/0378-1119(91)90420-g.
The thrombin inhibitor, hirudin, from the leech Hirudo Medicinalis, is the most powerful natural anticoagulant known. It has been characterized as a polypeptide of 65 amino acids which exhibits its anticoagulant properties by binding tightly and specifically to alpha-thrombin. The potency and specificity of hirudin have generated interest on its possible use in the treatment or prophylaxis of various thrombotic diseases. We have used the baculovirus expression system to efficiently produce active hirudins in insect cells. The Autographa californica nuclear polyhedrosis virus has proved useful as a helper-independent viral expression vector for high-level production of recombinant proteins in cultured insect cells. Hirudin variants (HV1 and HV2) were produced in infected insect cells as secreted proteins by joining their coding sequences to the leader peptide sequence of the vescicular stomatitis virus G protein. The recombinant products were biologically active and, interestingly, N-terminal sequencing of HV1 revealed that the heterologous leader peptide is correctly removed.
来自医用水蛭（Hirudo Medicinalis）的凝血酶抑制剂水蛭素，是已知最强效的天然抗凝剂。它被鉴定为一种由65个氨基酸组成的多肽，通过与α-凝血酶紧密且特异性地结合来展现其抗凝特性。水蛭素的效力和特异性引发了人们对其在治疗或预防各种血栓性疾病中可能用途的兴趣。我们利用杆状病毒表达系统在昆虫细胞中高效生产活性水蛭素。苜蓿银纹夜蛾核型多角体病毒已被证明是一种有用的无需辅助病毒的病毒表达载体，可用于在培养的昆虫细胞中高水平生产重组蛋白。水蛭素变体（HV1和HV2）在感染的昆虫细胞中作为分泌蛋白产生，方法是将它们的编码序列与水泡性口炎病毒G蛋白的前导肽序列连接。重组产物具有生物活性，有趣的是，HV1的N端测序显示异源前导肽被正确切除。
