猪囊尾蚴囊液16kDa特异性蛋白鉴定及其N-末端氨基酸序列测定

IDENTIFICATION OF A 16 KDA SPECIFIC PROTEIN FROM CYST FLUID OF CYSTICERCUS AND ITS N-TERMINAL AMINO ACID SEQUENCING

为了寻找猪囊尾蚴病诊断生物标记物，本研究从自然感染猪体肌肉分离、收集囊尾蚴，抽取囊液制备粗抗原，用其免疫家兔获得抗血清，再用SDS-PAGE分析囊尾蚴囊液总蛋白。以特异性抗血清通过Western Blot鉴定低分子量高特异性蛋白，测定其N-末端氨基酸序列并与相关资料进行对比分析，初步确定所鉴定蛋白归属。研究结果表明，从囊尾蚴囊液中鉴定出一低分子量为16kDa的特异性蛋白，其N-末端起始的10个氨基酸序列为DLSKGEWQLV，该序列与猪囊尾蚴囊液肌红蛋白的同源性为80％。鉴于其对抗猪囊尾蚴血清具有高特异性和高反应性，可将其作为生物标记物，用于猪囊虫病的免疫学诊断研究。

In order to search a molecular biomarker with low molecular weight for the diagnosis of cysticercosis, cysticercuses from the muscle of a naturally infected pig were isolated and collected. Rough antigen was prepared from the cyst fluid of the cysticercus and used to immunize rabbits. General protein of the cyst fluid was analyzed by SDS-PAGE. Specific protein was identified with Western Blot method by antiserum obtained from the immunized rabbits. N-terminal amino acids of the identified protein were detected and sequenced with Edman degradation. The results demonstrated that a 16 kDa protein with high specificity was identified from the cyst fluid of cysticercus and 10 amino acids were detected in its N-terminal and sequenced as DLSKGEWQLV. It is shown that the 16 kDa protein has 80% identity to myoglobin. It will be useful in the further study on the immunodiagnosis of cysticercosis.