蚯蚓纤溶酶基因P_(239)的原核表达、纯化及活性测定

Gene Expression of Earthworm Fibrinolytic Enzyme in E.coli, Purification and Bioassay

通过RT PCR方法从蚯蚓 (Lumbricusbimastus)中获得蚯蚓纤溶酶基因 ,命名为P2 3 9,含有 85 2个核苷酸 ,成熟肽编码 2 3 9个氨基酸 ,通过Genbank序列同源性比较 ,与已知的蚯蚓纤溶酶有一定的同源性 ,为首次获得的新基因。随后构建了 pBV2 2 0 /P2 3 9重组质粒 ,在大肠杆菌DH5α中进行原核表达 ,再经亲和层析柱纯化复性 ,其产物经活性测定 ,确定不仅具有激酶作用 ,而且具有直接溶栓活性。

The gene of earthworm fibrinolytic enzyme from earthworm(Lumbrukinase bimastus) was obtained by RT-PCR, named P-{239},The gene consists of 852 nucleotides that code 239 amino acids. Nucleotides order compared in Genbank, share certain homology with those known Lumbrukinase,it is a new gene,P-{239} was expressed in pBV220 expression system as inclusive body protein. Then the protein was purified by G25 and Sephrose 4B.The protein shows not only fibronolytic activity,but also directly fibrous protein dissolving activity.