細(xì)粒棘球絳蟲抗原B8-KDa亞單位在蟲體發(fā)育過程中的選擇性表達(dá)
發(fā)布時(shí)間：2018-12-29 14:09
【摘要】： 目的:研究細(xì)粒棘球絳蟲抗原B (EgAgB) 8-kDa亞單位基因家族成員(EgAgB8/1、EgAgB8/2、EgAgB8/3、EgAgB8/4、EgAgB8/5)在蟲體發(fā)育各階段(蟲卵、棘球蚴生發(fā)層、原頭蚴及成蟲)基因表達(dá)的差異,為包蟲感染中間宿主和終末宿主的免疫學(xué)診斷篩選特異性優(yōu)勢(shì)表達(dá)抗原。方法:分別從細(xì)粒棘球絳蟲成蟲、蟲卵、原頭蚴和棘球蚴生發(fā)層提取總RNA,用AMV First Strand cDNA Synthesis Kit反轉(zhuǎn)錄成cDNA,根據(jù)EgAgB 5個(gè)亞單位(EgAgB8/1、EgAgB8/2、EgAgB8/3、EgAgB8/4、EgAgB8/5)序列設(shè)計(jì)跨越內(nèi)含子的基因特異性引物;基因表達(dá)差異應(yīng)用SYBR Green I熒光實(shí)時(shí)定量PCR(Real-time PCR)技術(shù)進(jìn)行定量分析并用內(nèi)參對(duì)照基因actin II來標(biāo)準(zhǔn)化定量結(jié)果。結(jié)果:成功建立標(biāo)準(zhǔn)曲線和SYBR Green I熒光實(shí)時(shí)定量PCR(Real-time PCR)技術(shù)對(duì)EgAgB階段性基因表達(dá)的定量分析方法;分析顯示,EgAgB8/1和EgAgB8/2在棘球蚴生發(fā)層大量表達(dá)(68.874和16.258), EgAgB8/3和EgAgB8/5在成蟲階段有大量表達(dá)(1905.512 and 180.315),EgAgB8/4在成蟲、蟲卵和原頭蚴的表達(dá)水平(0.001、0.088和0.102)均較棘球蚴生發(fā)層表達(dá)量(3.576)低。結(jié)論:EgAgB 5個(gè)亞單位的基因表達(dá)在蟲體發(fā)育各個(gè)階段顯現(xiàn)出明顯差異:①EgAgB8/1和EgAgB8/2可作為包蟲感染中間宿主免疫學(xué)診斷最佳候選目的抗原,EgAgB8/3可作為包蟲感染終末宿主免疫學(xué)診斷的靶抗原;②EgAgB 5個(gè)亞單位的階段差異表達(dá)可能與其蟲體發(fā)育不同階段特有的生物學(xué)功能有關(guān),有待進(jìn)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.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R392
本文編號(hào)：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.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R392
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
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