华支睾吸虫抗原CsSQ1和CsSQ2的免疫学虫体定位  

Immunolocalization of antigens CsSQ1 and CsSQ2 of Clonorchis sinensis

作　　者：孙青松 于妮娜 尚信池 钱瑞程 李国江 刘明远 

机构地区：[1]吉林农业科技学院动物科技学院预防兽医学吉林省重点实验室,吉林吉林132000 [2]吉林大学动物医学学院人兽共患病研究教育部重点实验室,吉林长春130062

出　　处：《中国兽医科学》2017年第10期1299-1302,共4页Chinese Veterinary Science

基　　金：吉林市科技发展计划资助项目(20166034);吉林农业科技学院博士启动基金项目(吉农院合字2015第243号);吉林农业科技学院种子基金项目(吉农院合字2016第Z01号);吉林农业科技学院校级重点学科培育项目(吉农合字2015第X055号)

摘　　要：为了对华支睾吸虫重组抗原的特性进行鉴定及虫体定位,进而推测其生物学功能,通过对囊蚴与成虫的cDNA表达文库的免疫学筛选,获得若干个强反应原性抗原蛋白,在此基础上,发现2个含有20~30个氨基酸的高度重复序列CsSQ1和CsSQ2,利用基因序列合成多肽,再将多肽与载体蛋白进行偶联,制备重组抗原rCsSQ1和rCsSQ2,免疫动物制备多克隆抗体,利用硫酸铵沉淀法及亲和层析法纯化抗体,最后利用免疫荧光技术对重组抗原进行免疫学虫体定位。结果显示,重组抗原rCsSQ1和rCsSQ2分别定位于华支睾吸虫的口吸盘和腹吸盘。利用免疫荧光技术成功地确定了华支睾吸虫这两种抗原在虫体中的定位,推测它们可能与华支睾吸虫的免疫黏附作用有关,为华支睾吸虫抗原的免疫学特性及其在诊断中的应用研究提供了依据。To study the localization and characteristics of the recombinant antigens of Clonorchis sinensis,two amino acids highly repetitive sequences were obtained by construction and screening of a cDNA expression libraries of C.sinensis etacercaria and adult worms.Then we synthesized peptides with highly repetitive gene sequences and coupling with the carrier protein. Animals were immunized and sera was collected.Then antibodies were purified by ammonium sulfate precipitation and affinity chromatography. At last antigenic genes were located by immunofluorescence technique.In results,recombinant antigens CsSQ1 and CsSQ2 of C.sinensis mainly distributed in oral sucker and ventral suckers,respectively.The localization of the recombinant antigens of C.sinensis was confirmed.We speculate that the recombinant antigens may be related to the immune adhesion of C.sinensis.It provided the reference for the applied research in diagnosis and immunological characteristics of antigens of C.sinensis in the future.

关 键 词：华支睾吸虫 重组抗原 特性 免疫学定位 

分 类 号：S852.735[农业科学—基础兽医学]