东方田鼠天然抗日本血吸虫感染相关蛋白质的分离和纯化

目的从东方田鼠血清中分离纯化与其天然抗日本血吸虫感染特性相关的蛋白质。方法以分子筛色谱及高效液相色谱技术对东方田鼠血清蛋白质进行分离和纯化,以体外日本血吸虫童虫杀伤实验确定具有抗日本血吸虫童虫活性蛋白质,并对活性蛋白质进行N端氨基酸序列测定,通过检索蛋白质数据库以确定活性蛋白质的性质和种类。结果利用分子筛色谱及高效液相色谱技术从东方田鼠血清中获得了三个对日本血吸虫童虫具有明显杀伤活性的蛋白质组份1-1、4-1、6-3,经对4-1的N末端氨基酸序列进行测定,其序列为DAHKSEIAHR,检索蛋白质数据库,该蛋白质为白蛋白样蛋白质,但与人或其它动物的白蛋白进行比较,存在较大程度的氨基酸残基变。结论东方田鼠血清中的白蛋白是东方田鼠天然抗日本血吸虫感染的重要效应分子。

Isolation and purification of proteins associated with natural resistance to Schistosoma japonicum from Microtus fortis serum

The isolation and purification of serum proteins which associated with natural resistance to S.japonicum in M.fortis serum was carried out by means of Molecular sieve chromatography and High Performance Liquid Chromatography,the amino acid sequence of active protein which the killing activity to S.japonicum schistosomula was confirmed in vitro was detemined by Edman degradation technique,and BLAST was used to identify the protein.Three active protein components(1-1,4-1,6-3)which showed significant killing effect to S.japonicum schistosomula in vitro were gained,the mortality rate of S.japonicum schistosomula was 12.2%,11.7% and 20.1%,respectivelly(P<0.05).The N-terminal amino acid sequence of 4-1 is DAHKSEIAHR,BLAST showed that this is an albumin-like protein,but some mutations in amino acid sequence were observed when compared with the albumin derived from other species.All these date suggests that albumin of M.fortis is an effective molecule which associated with the characteristic of natural resistance to schistosomiasis japonica.