Ge Tao, Sun Zhao-Jun, Fu Shi-Hong, Liang Guo-Dong
Institute for virus disease control and prevention, No.100 YingXin Jie, XuanWu Qu, Beijing, China.
Protein Expr Purif. 2005 Jul;42(1):20-8. doi: 10.1016/j.pep.2005.04.005.
Lumbrokinase (PI239, GenBank Accession No. AF433650) from the earthworm Lumbricus bimastus has been identified. The cDNA of PI239 is composed of 852bp and includes an open reading frame that encodes two parts of the protein: a signal peptide of 44 amino acids and a mature peptide of 239 residues. The cDNA of PI239 exhibits a high degree of sequence identity with other lumbrokinase genes, ranging from 87.6% (F-III-I) to 98.3% (EFE-3). The gene encoding the native form of PI239, with a 5' non-functional end removed, was obtained by PCR amplification and was sub-cloned into pPICZalpha-A, a yeast expression and secretion vector. SDS-PAGE and Western blot analyses showed that rPI239 secreted into the culture medium was specifically recognized by the wild type lumbrokinase polyclonal antibody and was able to dissolve artificial fibrin plates.
已鉴定出来自双胸蚓的蚓激酶（PI239，GenBank登录号AF433650）。PI239的cDNA由852bp组成，包含一个开放阅读框，该阅读框编码蛋白质的两个部分：一个44个氨基酸的信号肽和一个239个残基的成熟肽。PI239的cDNA与其他蚓激酶基因表现出高度的序列同一性，范围从87.6%（F-III-I）到98.3%（EFE-3）。通过PCR扩增获得了编码去除了5'非功能末端的天然形式PI239的基因，并将其亚克隆到酵母表达和分泌载体pPICZalpha-A中。SDS-PAGE和蛋白质免疫印迹分析表明，分泌到培养基中的重组PI239能被野生型蚓激酶多克隆抗体特异性识别，并且能够溶解人工纤维蛋白平板。
