| 细粒棘球蚴特异性抗原基因P-29的原核表达及免疫学鉴定 |
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| 引用本文： | 李洁,王志钢,郝喜燕,陈献威,王红霞,李树裕,杨娇馥,杨军. 细粒棘球蚴特异性抗原基因P-29的原核表达及免疫学鉴定[J]. 中国寄生虫学与寄生虫病杂志, 2010, 28(2): 125-128 |
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| 作者姓名： | 李洁 王志钢 郝喜燕 陈献威 王红霞 李树裕 杨娇馥 杨军 |
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| 作者单位： | 1内蒙古大学生命科学学院，哺乳动物生殖生物学与生物技术教育部重点实验室，呼和浩特 010021；2 内蒙古自治区呼和浩特市环境科学研究所，呼和浩特 010030 |
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| 基金项目： | 国家基础科学人才培养基金(No.J0730648)~~ |
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| 摘 要： | 目的对细粒棘球蚴P-29基因进行克隆、表达和免疫反应性分析。方法以细粒棘球蚴总RNA为模板,反转录PCR扩增P-29基因,将其克隆至原核表达载体pET44a(+)中,构建重组原核表达载体pET-P-29,转入大肠埃希菌BL21(DE3)中,用异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)观察重组蛋白的表达情况,用蛋白纯化试剂盒纯化蛋白,蛋白质印迹(Western blotting)分析重组蛋白与细粒棘球蚴病患者血清的免疫反应性。结果PCR、双酶切和DNA测序结果表明,重组质粒pET-P-29构建成功。SDS-PAGE结果显示,重组蛋白Nus-P-29的相对分子质量(Mr)约为93000,纯化后的蛋白浓度为0.78mg/ml。重组蛋白Nus-P-29能被细粒棘球蚴病患者血清识别。结论细粒棘球蚴P-29基因表达成功,纯化后的重组蛋白Nus-P-29具有较强的免疫反应性。
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| 关 键 词： | 细粒棘球蚴；特异性抗原基因；原核表达 |
Prokaryotic Expression and Immunologic Identification of P-29 Gene from Echinococcus granulosus Hydatid Cyst |
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| LI Jie,WANG Zhi-gang,HAO Xi-yan,CHEN Xian-wei,WANG Hong-xia,LI Shu-yu,YANG Jiao-fu,YANG Jun. Prokaryotic Expression and Immunologic Identification of P-29 Gene from Echinococcus granulosus Hydatid Cyst[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2010, 28(2): 125-128 |
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| Authors: | LI Jie WANG Zhi-gang HAO Xi-yan CHEN Xian-wei WANG Hong-xia LI Shu-yu YANG Jiao-fu YANG Jun |
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| Affiliation: | LI Jie1,WANG Zhi-gang1,HAO Xi-yan1,CHEN Xian-wei1,WANG Hong-xia1,LI Shu-yu1,YANG Jiao-fu1,YANG Jun2(1 College of Life Science,Inner Mongolia University,Key Laboratory of Mammal Reproductive Biology , Biotechnology,Ministry of Education,Hohhot 010021,China,2 Hohhot Institute of Environmental Sciences,Hohhot 010030,China) |
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| Abstract: | Objective To clone and express P-29 gene of Echinococcus granulosus, and analyze its immunore- activity. Methods Total RNA was extracted from the hydatid cyst of E. granulosus and its P-29 gene was amplified by RT-PCR. The PCR product was cloned into pET44a(+) vector. The recombinant plasmid was transformed into E. coli BL21 (DE3) and followed by expression of the protein induced by IPTG. The protein was purified, and tested by SDS- PAGE. Its immunoreactivity was examined by Western blotting. Results The re... |
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| Keywords: | Echinococcus granulosus Specific antigen gene Prokaryotic expression |
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