MDV超强毒株GX0101感染鸡脾的蛋白质组学研究

发布时间：2019-02-24 17:34

【摘要】：MD是由MDV感染引发的一种常见的恶性淋巴细胞瘤传染病。为进一步研究MDV的致瘤机制,本研究通过MDV超强毒株GX0101感染SPF鸡群构建动物模型:(1)将1日龄的SPF雏鸡分为试验组和对照组各40只,试验组每只雏鸡腹腔注射200μL的GX0101型MDV毒株,对照组每只雏鸡等量注射CEF原代细胞液体。此后在第7 d、14 d、21 d、30 d、45 d和60 d各采取3只试验组和对照组鸡的心、肝、脾、肺、肾、腺胃、胸腺,脑,胰腺等组织样品和血液,通过荧光定量PCR检测不同时间点血液中meq和gB的表达量,通过HE染色方法观察各组织病变情况;(2)通过Label-free技术分析了45 d后试验组和对照组鸡脾脏的蛋白质组学,并对差异蛋白进行了GO分析和KEGG分析;(3)通过荧光定量PCR研究了注射GX0101毒株和CEF细胞后不同时间点试验组和对照组鸡各组织的PRNP相对表达量。通过本研究得出结论:(1)荧光定量PCR检测发现,MDV标志性基因meq和gB在注射GX0101后45 d表达量最高,并多个组织在45 d观察到了肿瘤,meq和gB表达量与肿瘤的形成相关;(2)试验组同对照组相比,其中有690个蛋白表达上调,390个蛋白下调。GO分析注释结果显示,这些差异蛋白涉及到的生物过程主要是代谢过程,转运和氧化还原过程。KEGG经典通路分析显示这些差异蛋白主要涉及到癌症相关的通路,朊蛋白相关的通路,信号通路以及其它通路等。(3)PRNP基因表达量与MDV感染具有相关性,并测得PRNP基因在试验组鸡群多个组织中的表达量基本高于对照组鸡群。PRNP基因在注射组多个组织的表达量随着时间的变化而变动,在45 d达到峰值,此后逐渐下降至60d并处于较低的转录水平。
[Abstract]:MD is a common malignant lymphocytoma infection caused by MDV infection. In order to further study the tumorigenic mechanism of MDV, we established an animal model of SPF chickens infected with MDV supervirulent GX0101 strain. (1) the 1-day-old SPF chicks were divided into two groups: the experimental group and the control group (n = 40). In the experimental group, 200 渭 L GX0101 type MDV strain was injected intraperitoneally in each chick, while in the control group, the primary cell fluid of CEF was injected into the same quantity. After that, the heart, liver, spleen, lung, kidney, glandular stomach, thymus, brain, pancreas and other tissue samples and blood of 3 chickens in the experimental group and the control group were taken at the 7th day, 14 days, 21 days, 30 days, 45 days and 60 days, respectively. The expression of meq and gB in blood at different time points were detected by fluorescence quantitative PCR, and the pathological changes of tissues were observed by HE staining. (2) the proteomics of spleen of the experimental group and the control group were analyzed by Label-free technique after 45 days, and the differential proteins were analyzed by GO and KEGG. (3) the relative expression of PRNP in chicken tissues at different time points after injection of GX0101 and CEF cells was studied by fluorescence quantitative PCR. The results of this study were as follows: (1) fluorescence quantitative PCR analysis showed that the expression of MDV signature genes meq and gB was the highest at 45 days after GX0101 injection, and the tumor was observed in several tissues at 45 days. The expression of meq and gB was related to the formation of tumor. (2) compared with the control group, 690 proteins were up-regulated and 390 proteins down-regulated in the experimental group. The results of GO analysis showed that the biological processes involved in these differential proteins were mainly metabolic processes. Transport and redox processes. KEGG classic pathway analysis shows that these differential proteins are mainly involved in cancer-related pathways, prion protein-related pathways, (3) the expression of PRNP gene was correlated with MDV infection. The expression of PRNP gene in multiple tissues of experimental group was higher than that in control group. The expression of PRNP gene in multiple tissues of injection group changed with time and reached its peak at 45 days. After that, it gradually decreased to 60 days and was at a lower level of transcription.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.31

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