草原兔尾鼠卵透明带3蛋白的免疫原性及其抗血清对体外精卵结合的影响  

Immunogenicity of recombinant Lugurus zone pellucida-3 expressed in E.coli and efficacy of its antiserum to affect in vitro mouse sperm-egg binding in vitro

作　　者：张爱莲[1] 庄淑珍[1] 刘菲[1] 张富春[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《中国免疫学杂志》2008年第3期244-247,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(No30360062);新疆教育厅创新群体项目(No:XJEDU2004G02);新疆大学青年教师科研启动金(NoQN040119)资助项目

摘　　要：目的:大肠杆菌表达的重组草原兔尾鼠透明带3(Recombinant Luguruszone pellucida3,r-LZP3)蛋白的免疫原性直接影响抗体的滴度,通过对r-LZP3免疫原性的分析,获得特异性的抗血清,并探讨r-LZP3和抗血清对体外精卵结合的影响。方法:在大肠杆菌中以包涵体形式表达r-LZP3,经过变性、复性及纯化后作为抗原,用LZP3DNA疫苗初免小鼠后,再用r-LZP3进行免疫加强,产生特异性的LZP3抗血清;通过EL1SA测定抗体应答的水平;蛋白印迹、间接免疫荧光和免疫组化方法检测抗血清同r-LZP3、鼠卵母细胞表面天然ZP的反应性;精卵结合实验观察r-LZP3、抗血清体外对鼠精卵结合能力的影响。结果:用LZP3DNA疫苗免疫制备的抗血清在免疫鼠中产生了较高的抗体滴度,并且可以识别大肠杆菌表达的r-LZP3以及鼠卵细胞表面天然的ZP,抗血清和r-LZP3也都能在体外抑制鼠的精卵结合。结论:大肠杆菌表达的r-LZP3有较好的免疫原性,r-LZP3和抗血清能够在体外阻止鼠的精卵结合。Objective：Present study was aimed to analyze the immunogenicity of recombinant Lugurus zona pellucida-3 protein （r- LZP3） expressed in E. coli, and to evaluate the efficacy of its antiserum to affect in vitro mouse sperm-egg binding. Methods： Female mouse were immunized using LZP3 DNA vaccine for priming, and boosted with r-LZP3 protein. The antibody response level of LZP3 antisera was determined by ELISA. The immunoreactivity and specificity of the anti-LZP3 antisera were tested by immunoblot, indirect immunofluorescence and immunobistochemistry using native mouse ZP. A swam-zone pellucida binding assay was used to evaluate the efficacy of anti-LZP3 antisera to inbibit mouse sperm-egg binding in vitro.Results： The result of ELISA showed that LZP3 DNA vaccine was able to induce higher antibody titers in mouse.Antiserum could specifically recognize or bind to r-LZP3 protein expressed in E. coli and native mouse ZP in vitro.The LZP3 antisera and r-LZP3 protein also inhibited sperm-egg binding in vitro. Conclusion：These results show that r-LZP3 protein is of strong immunogericity.

关 键 词：草原兔尾鼠卵透明带3 免疫原性 抗血清 卵母细胞 精卵结合 

分 类 号：R392.11[医药卫生—免疫学]