4B4基因在褐飞虱不同发育阶段表达分析.docVIP

4B4基因在褐飞虱不同发育阶段的表达分析 黎璐 广州大学生命科学学院 摘要 本实验以探究4B4基因在褐飞虱不同发育阶段的表达是否存在差异为最终目的，通过对1至4龄的褐飞虱虫体总RNA的抽提、设计4B4基因的特异性引物进行RT-PCR，结合琼脂糖凝胶电泳等系列实验，初步证实褐飞虱在其个体发育的不同阶段4B4基因的表达确实存在一定的差异。由此反映出基因的表达调控具有时空特性，在个体发育的进程中，某些基因的表达水平是受到与之平行或相关的调节的。对4B4基因随时间推移的个体表达情况作进一步的分析，将有贡献于水稻和褐飞虱的互作研究，为褐飞虱及相类似的水稻害虫的生物防治提供可参考的实验依据。 关键词 褐飞虱 基因表达 RNA RT-PCR 琼脂糖凝胶电泳 4B4 gene expression analysis in different developmental stages of the brown planthopper LiLu School of Life Sciences，Guangzhou University ABSTRACT The ultimate goal In this experiment is to explore the expression of 4B4 gene in different developmental stages of the brown planthopper to see whether differences exist .By extracting the total RNA of 4 kinds of brown planthopper in different life stages , designing 4B4 gene-specific primers for RT-PCR, combined with agar agarose gel electrophoresis and other series of experiments,it’s initially confirmed 4B4 gene expression does exist some differences of the brown planthopper in its different stages of ontogeny. Thus reflecting the regulation of gene expression with spatial and temporal characteristics, in the process of ontogeny, the expression levels of certain genes is subject to parallel or related regulation. Further analysis of 4B4 gene expression for individuals over time will contribute to the studies about the interaction between brown planthopper with rice,and provide an experimental basis for the reference to the brown planthopper and similar biological control of insects of rice. KEY WORDS Niaparvata lugens Stal; Gene Expression; RNA; RT-PCR; Agarose Gel Electrophoresis 目录 前言 …………………………………………………………………………………1 材料与方法 ………………………………………………………………………5 2.1 实验材料、器材和试剂 ……………………………………………………5 2.1.1实验材料 …………………………………………………………………5 2.1.2实验器材 ………………………………………………………………… 5 2.1.3实验试剂 …………………………………………………………………5 2.2 实验方法 …………………………………………………………………… 5 2.2.1试剂配制 ………………………………………………………………… 5 2.2.2设计引物 ………………………………………………………………… 6 2.2.3 RNA制备 …………………………………………………………………7 2.2.4 RT-PCR反应