目录
摘要Ⅱ
关键词Ⅱ
AbstractⅢ
引言
引言1
1 材料与方法1
1.1材料与仪器 1
1.1.1实验材料 1
1.1.2实验试剂及耗材1
1.2实验方法2
1.2.1过表达载体的构建2
1.2.2BMDC细胞体外诱导培养5
1.2.3流式细胞检测6
2结果与分析7
2.1过表达载体构建结果4
2.1.1目的基因PCR结果6
2.1.2菌落PCR电泳结果 8
2.1.3测序结果8
2.2流式细胞仪检测结果 9
2.2.1过表达miRNA对树突状细胞表型的影响9
2.2.2 抑制miRNA片段对树突状细胞表型的影响10
2.3靶基因信号通路预测结果11
2.3.1miR19a的靶基因及信号通路预测11
2.3.2 miR2131的靶基因及信号通路预测15
3讨论21
致谢 21
参考文献21
鸡miRNA19a与miRNA2131对鸡树突状细胞免疫应答的影响
摘 要
禽传染性支气管病毒（Avian infectious bronchitis virus ，IBV）是一种引起禽呼吸道疾病的一种冠状病毒；树突状细胞（Dendritic cell， DCs）是机体中最重要的专职抗原提呈细胞，其对病毒的识别和递呈直接决定了对病毒的清除效果。近期研究发现非编码RNA广泛参与了宿主对病毒入侵的调控。本研究前期研究结果发现IBV入侵DC细胞之后，mir19a与mir2131表达量显著下调。为进一步探究mir19a与mir2131对DCs免疫功能的影响，我们首先扩增构建mir19a与mir2131的过表达载体，研究这两个microRNA过表达对鸡骨髓源树突状细胞的影响；其次干扰DCs中的mir19a与mir2131检测其对DCs表型的影响。再次，本研究通过生物信息学的方法预测了mir19a与mir2131的靶基因及其相关信号通路，结果显示 *51今日免费论文网|www.51jrft.com +Q: ￥351916072$ 
mir19a的靶基因主要参与了肌动蛋白骨架的调节、钙离子信号通路、局灶性沾连、间隙连接和MAPK信号通路有关。与之不同mir2131的靶基因则主要参与了局灶性沾连、ErbB 信号通路、mTOR 信号通路、FoxO 信号通路、间隙连接、Toll样受体信号通路、NOD样受体信号通路和MAPK信号通路有关，DC细胞功能是吞噬抗原之后与T细胞粘连将信号传达给T细胞，其中过程包括细胞的识别、细胞间连接、细胞的存活、细胞的迁移、水解，正与mir19a和mir2131调节的信号通路有关，与本实验研究目的相符合
EFFECTS OF CHICKEN MIRNA19A AND MIRNA2131 ON IMMUNE RESPONSE OF CHICKEN DENDRITIC CELLS
ABSTRACT
Avian infectious bronchial virus (IBV) is a coronavirus that causes respiratory diseases in birds; DC cells are antigenpresenting cells, which are responsible for transmitting viral information to T cells and play an important role after virus invasion. Recent studies have found that noncoding RNA has an effect on the immune response of virus invasion; previous studies have found that after IBV invades DC cells, the expression levels of mir19a and mir2131 are downregulated in order to explore the effects of mir19a and mir2131 on DC The influence of cells, we interfered and overexpressed mir19a and mir2131, transfected into chicken bone marrow dendritic cells (BMDCs), and detected by flow cytometry, found that mir19a and mir2131 were DC cell maturation has an effect. The bioinformatics method was used to predict the signal pathways of mir19a and mir2131 target genes. The results showed that mir19a and Actin skeleton regulation, Calcium signal pathway, Focal adhesion, Gap junction and MAPK Signaling pathway, mir2131 is related to Focal adhesions, FoxO signaling pathway, mTOR signaling pathway,ErbB signaling pathway , Tolllike receptor signaling ,Gap junction, pathway, NODlike receptor signaling pathway It is related to the MAPK signaling pathway. The function of DC cells is to adhere to T cells after phagocytosis of antigens and transmit signals to T cells. The process includes cell recognition, cell connection, cell survival, cell migration, and hydrolysis. It is related to mir19a It is related to the signal pathway regulated by mir2131, which is consistent with the purpose of this experiment.

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