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  

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

作　　者：邬国军[1] 胡维新[1] 秦志强[1] 罗赛群[1] 俞远京[2] 王勇[3] 彭兴华[1] 

机构地区：[1]中南大学生物科学与技术学院分子生物研究中心 [2]中南大学实验动物学部 [3]中国科学院亚热带农业生态研究所(原中国科学院长沙农业现代化研究所)

出　　处：《中国人兽共患病杂志》2005年第8期688-692,共5页Chinese Journal of Zoonoses

基　　金：国家自然科学基金(批准号:30070403);国家科技部"九五"科技攻关项目(编号:96-A23-06-04);湖南省科技厅"九五"科技攻关配套基金项目资助。

摘　　要：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。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 sehistosomula was corfirmed in vitro was determined 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 sehistosomissis japonica.

关 键 词：东方田鼠 日本血吸虫 天然抗性 感染 

分 类 号：R383.2[医药卫生—医学寄生虫学]