含弓形虫ROP1基因真核表达重组质粒DNA免疫小鼠的研究Ⅲ.免疫鼠肌组织表达产物免疫酶法定位及血清IgG抗体测定  

VACCINATING MICE WITH EUKARYOTIC EXPRESSION RECOMBINANT PLASMID DNA CODING RHOPTRY PROTEIN 1 GENE FROM TOXOPLASMA GONDIIⅢ.Detecting the Expression Product in Muscle Tissue and IgG Antiboes in Serum of the Immunized Mice

作　　者：郭虹[1] 陈观今[1] 郑焕钦[1] 周永安[1] 

机构地区：[1]中山医科大学寄生虫学研究所,广州510089

出　　处：《中国人兽共患病杂志》1999年第6期11-13,共3页Chinese Journal of Zoonoses

基　　金：中国博士点学科科研基金

摘　　要：目的　用所构建的弓形虫ｐｃＤＮＡ３－ＲＯＰ１ 真核表达重组质粒，经肌肉注射免疫小鼠，观察它在肌组织中的表达及不同免疫途径所诱导的体液免疫应答。方法　碱裂解法大量制备ｐｃＤＮＡ３－ＲＯＰ１ 质粒，免疫ＢＡＬＢ／ｃ小鼠，每只鼠注射１００ｕｇ，两周后同量加强免疫一次，以ｐｃＤＮＡ３ 空质粒及生理盐水组为对照。于免疫后第５０ 天用间接免疫酶法检测注射局部肌组织重组蛋白的表达；ＥＬＩＳＡ法测定ＩｇＧ抗体滴度。结果　免疫鼠肌组织石蜡切片呈特异性阳性反应；血清ＩｇＧ抗体９０天后测定为阳性；皮下及肌肉不同免疫途径血清均显示阳性结果，无显著性差异。结论　ｐｃＤＮＡ３－ＲＯＰ１质粒ＤＮＡ 免疫小鼠后，肌组织内有重组蛋白表达。Aim\ DNA vaccinating BALB/c mice with the eukaryotic expression recombinant plasmid,pcDNA3,containing ROP1 gene from Toxoplasma gondii to observe the expression product in the muscle tissue and the titer of IgG antibody in the immunized mice.Methods\ Large scale preparation of plasmid DNA by alkaline lysis,the DNA were injected through quadriceps muscles of left leg each mouse at the dosage of 100μg.A booster vaccine was given at the same dosage after two weeks.Control groups were injected with pcDNA3 blank plasmid and normal saline respectively.After 50 days of the booster injection,the expression protein was detected with indirect immunohistochemistry assay and the titer of IgG antibody was detected by ELISA.Results\ The positive result of the indirect immunohistochemistry assay was showed in immunal group.The titer of IgG antibody was potitive in both vaccinated groups,which injected by muscle and by skin after 90 days.Conclusion\ After vaccinating BALB/c mice with the recombinant plasmid,pcDNA3 ROP1,by injection muscle directly,the recombinant protein was expressed in the muscle tissue and elicited the humoral immuno response in mice.

关 键 词：弓形生 棒状体蛋白 重组质粒 DNA免疫 小鼠 

分 类 号：R382.5[医药卫生—医学寄生虫学] R531.8[医药卫生—基础医学]