Zhang Jinhua, Meng Wenli, Wang Chunhua, Wu Zhiqiang, Wu Guotu, Xu Yunlu
School of Pharmacy, Fujian Medical University, Fuzhou 350004, PR China; Department of Pharmacy, Fujian Medical University Union Hospital, Fuzhou 350001, PR China.
Protein Expr Purif. 2013 Sep;91(1):85-90. doi: 10.1016/j.pep.2013.07.009. Epub 2013 Jul 23.
The plasminogen activator (PA) in snake venom, a serine protease, can convert plasminogen to active plasmin, indirectly causing the degradation of fibrin. It is difficult to purify sufficient snake venom PA (SV-PA) for clinical applications due to the low SV-PA content in venom. The gene encoding PA was obtained from the venom gland of Gloydius brevicaudus using RT-PCR with primers designed according to the N-terminal amino acids of G. brevicaudus venom PA (GBV-PA), was cloned into the prokaryotic expression vector pET-42a, and recombinant GBV-PA (rGBV-PA) was expressed via Isopropyl-β-d-1-Thiogalactopyranoside (IPTG) induction. Like human tissue PA, the purified renatured rGBV-PA could significantly reduce the rabbit plasma euglobulin lysis time (ELT) and prevent thrombus formation in the inferior vena cava of rats. Within the dosage range, the dosage and effects were positively correlated.
蛇毒中的纤溶酶原激活剂（PA）是一种丝氨酸蛋白酶，可将纤溶酶原转化为活性纤溶酶，间接导致纤维蛋白降解。由于毒液中蛇毒PA（SV-PA）含量较低，难以纯化出足够用于临床应用的蛇毒PA。根据短尾蝮蛇毒PA（GBV-PA）的N端氨基酸设计引物，采用RT-PCR从短尾蝮蛇毒腺中获得编码PA的基因，将其克隆到原核表达载体pET-42a中，通过异丙基-β-D-硫代半乳糖苷（IPTG）诱导表达重组GBV-PA（rGBV-PA）。与人类组织PA一样，纯化复性后的rGBV-PA可显著缩短兔血浆优球蛋白溶解时间（ELT），并预防大鼠下腔静脉血栓形成。在剂量范围内，剂量与效果呈正相关。
