细粒棘球绦虫抗原B8-KDa亚单位在虫体发育过程中的选择性表达
发布时间：2018-12-29 14:09
【摘要】： 目的:研究细粒棘球绦虫抗原B (EgAgB) 8-kDa亚单位基因家族成员(EgAgB8/1、EgAgB8/2、EgAgB8/3、EgAgB8/4、EgAgB8/5)在虫体发育各阶段(虫卵、棘球蚴生发层、原头蚴及成虫)基因表达的差异,为包虫感染中间宿主和终末宿主的免疫学诊断筛选特异性优势表达抗原。方法:分别从细粒棘球绦虫成虫、虫卵、原头蚴和棘球蚴生发层提取总RNA,用AMV First Strand cDNA Synthesis Kit反转录成cDNA,根据EgAgB 5个亚单位(EgAgB8/1、EgAgB8/2、EgAgB8/3、EgAgB8/4、EgAgB8/5)序列设计跨越内含子的基因特异性引物;基因表达差异应用SYBR Green I荧光实时定量PCR(Real-time PCR)技术进行定量分析并用内参对照基因actin II来标准化定量结果。结果:成功建立标准曲线和SYBR Green I荧光实时定量PCR(Real-time PCR)技术对EgAgB阶段性基因表达的定量分析方法;分析显示,EgAgB8/1和EgAgB8/2在棘球蚴生发层大量表达(68.874和16.258), EgAgB8/3和EgAgB8/5在成虫阶段有大量表达(1905.512 and 180.315),EgAgB8/4在成虫、虫卵和原头蚴的表达水平(0.001、0.088和0.102)均较棘球蚴生发层表达量(3.576)低。结论:EgAgB 5个亚单位的基因表达在虫体发育各个阶段显现出明显差异:①EgAgB8/1和EgAgB8/2可作为包虫感染中间宿主免疫学诊断最佳候选目的抗原,EgAgB8/3可作为包虫感染终末宿主免疫学诊断的靶抗原;②EgAgB 5个亚单位的阶段差异表达可能与其虫体发育不同阶段特有的生物学功能有关,有待进一步研究。
[Abstract]:Objective: to study the members of the B (EgAgB) 8-kDa subunit gene family (EgAgB8/1,EgAgB8/2,EgAgB8/3,EgAgB8/4,EgAgB8/5) of Echinococcus granulosus antigen in the germinal layer of Echinococcus granulosus (Echinococcus granulosus). The difference of gene expression between protocercariae and adults is a specific and dominant antigen for immunological diagnosis of intermediate host and terminal host of hydatid infection. Methods: the total RNA, was extracted from the germinal layer of Echinococcus granulosus adult, egg, protocaria and echinococcus, respectively. The total RNA, was inverted by AMV First Strand cDNA Synthesis Kit and transcribed into cDNA, according to five EgAgB subunits (EgAgB8/1,EgAgB8/2,EgAgB8/3,EgAgB8/4,). EgAgB8/5) sequence to design gene specific primers across introns; The difference of gene expression was analyzed by SYBR Green I real-time quantitative PCR (Real-time PCR) and the quantitative results were standardized by internal control gene actin II. Results: the standard curve and SYBR Green I real-time quantitative PCR (Real-time PCR) were successfully established for the quantitative analysis of EgAgB gene expression. The results showed that EgAgB8/1 and EgAgB8/2 were highly expressed in germinal layer of echinococcus (68.874 and 16.258), EgAgB8/3 and EgAgB8/5 in adult stage (1905.512 and 180.315), EgAgB8/4 in adult. The expression levels of egg and protocercariae (0. 001, 0. 088 and 0.102) were lower than those of Echinococcus germinal layer (3.576). Conclusion: the gene expression of the five subunits of EgAgB is significantly different in all stages of body development: 1EgAgB8/1 and EgAgB8/2 can be used as the best candidate antigen for immunological diagnosis of hydatid infection. EgAgB8/3 can be used as a target antigen for immunological diagnosis of hydatid infection. The differential expression of the five subunits of 2EgAgB may be related to the specific biological function in different stages of the development of 2EgAgB, which needs further study.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
本文编号：2394929
[Abstract]:Objective: to study the members of the B (EgAgB) 8-kDa subunit gene family (EgAgB8/1,EgAgB8/2,EgAgB8/3,EgAgB8/4,EgAgB8/5) of Echinococcus granulosus antigen in the germinal layer of Echinococcus granulosus (Echinococcus granulosus). The difference of gene expression between protocercariae and adults is a specific and dominant antigen for immunological diagnosis of intermediate host and terminal host of hydatid infection. Methods: the total RNA, was extracted from the germinal layer of Echinococcus granulosus adult, egg, protocaria and echinococcus, respectively. The total RNA, was inverted by AMV First Strand cDNA Synthesis Kit and transcribed into cDNA, according to five EgAgB subunits (EgAgB8/1,EgAgB8/2,EgAgB8/3,EgAgB8/4,). EgAgB8/5) sequence to design gene specific primers across introns; The difference of gene expression was analyzed by SYBR Green I real-time quantitative PCR (Real-time PCR) and the quantitative results were standardized by internal control gene actin II. Results: the standard curve and SYBR Green I real-time quantitative PCR (Real-time PCR) were successfully established for the quantitative analysis of EgAgB gene expression. The results showed that EgAgB8/1 and EgAgB8/2 were highly expressed in germinal layer of echinococcus (68.874 and 16.258), EgAgB8/3 and EgAgB8/5 in adult stage (1905.512 and 180.315), EgAgB8/4 in adult. The expression levels of egg and protocercariae (0. 001, 0. 088 and 0.102) were lower than those of Echinococcus germinal layer (3.576). Conclusion: the gene expression of the five subunits of EgAgB is significantly different in all stages of body development: 1EgAgB8/1 and EgAgB8/2 can be used as the best candidate antigen for immunological diagnosis of hydatid infection. EgAgB8/3 can be used as a target antigen for immunological diagnosis of hydatid infection. The differential expression of the five subunits of 2EgAgB may be related to the specific biological function in different stages of the development of 2EgAgB, which needs further study.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R392
【参考文献】
相关期刊论文 前2条
1 史大中;中国囊性包虫病的地理分布[J];地方病通报;2000年01期
2 柴君杰;焦伟;伊斯拉音;常青;孟贺巴特;付承;韩玲玲;徐世东;孙立峰;白国琦;赛力克;马合木提;多鲁肯;薛海筹;冯正;;新疆北部地区囊型包虫病的流行现状[J];热带病与寄生虫学;2004年03期
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