摘   要核糖体（ribosome）是细胞中的一种细胞器，除哺乳动物成熟的红细胞外，细胞中都有核糖体存在。一般而言，原核细胞只有一种核糖体，而真核细胞具有两种核糖体（其中线粒体中的核糖体与细胞质核糖体不相同）。核糖体由大、小两个亚基（subunit）组成。由于沉降系数不同，核糖体又分为70S型和80S型两种。70S型核糖体主要存在于原核细胞的叶绿体、线粒体基质中，其小亚基单位为30S，大亚基单位为50S。80S型核糖体主要存在于真核细胞质中，其小亚基单位为40S，大亚基单位为60S。核糖体是细胞内一种核糖核蛋白颗粒(rib nucleoprotein particle），主要由RNA(rRNA）和蛋白质构成，之前一直认为其唯一功能是按照mRNA的指令将氨基酸合成蛋白质多肽链，所以核糖体是细胞内蛋白质合成的分子机器。近年来，已有报导证实了RPL10A参与植物对DNA病毒的防御。本课题采用病毒学诱导基因沉默技术（VIGS），在本氏烟草系统中研究拟南芥RPL10A同家族基因RPL7Ae的功能。研究表明，接种PVX/AtRPL7Ae重组病毒RNA后的本氏烟草出现了矮化，叶片卷缩等表型，同时也大大影响了植株果荚数量。由此可知，RPL7Ae基因在植物发育及生殖方面具有重要的功能。76636

Abstract Ribosomes (ribosome) is a cell organelle, in addition to the mature mammalian erythrocytes, the cells have ribosomes exist。 Generally, only one prokaryotic ribosome, and eukaryotic cells with two ribosomes (ribosomes which mitochondria with different cytoplasmic ribosomes)。 Ribosome by a large, two small subunit (subunit) components。 Since the sedimentation coefficient different type ribosomal 70S and 80S is pided into two kinds。 70S ribosome primarily in prokaryotic cells chloroplast, mitochondrial matrix, which is and 30S small subunit units, units of the large subunit 50S。 80S ribosome is mainly present in the eukaryotic cytoplasm, the unit is small subunit 40S, units of the large subunit 60S。 Ribosome is a ribonucleoprotein particle (ribonucleoprotein particle) cells, mainly composed of RNA (rRNA) and protein, until that its sole function is to follow the instructions mRNA amino acid polypeptide chain protein synthesis, the ribosome is so intracellular protein synthesis of molecular machines。 In recent years, it has been reported that RPL10A is involved in the defense of DNA virus。 In this study, the gene silencing technique (VIGS) was used to study the function of Arabidopsis RPL10A gene RPL7Ae in the tobacco system。 Studies have shown that after inoculation PVX/AtRPL7Ae recombinant virus RNA's tobacco the dwarfing, leaf curling phenotype。 At the same time, but also greatly affected the plant number of pods。 It can be known that the RPL7Ae gene has important function in plant development and reproduction。

毕业论文关键词：核糖体大亚基;土豆病毒X；病毒诱导基因沉默；拟南芥；本氏烟草

Key words: Ribosome large subunit；Potato virus X; VIGS；Arabidopsis thaliana; Nicotiana Benthamiana

目录 4

引言 5

1 材料与方法 6

1。1  实验材料以及设备 6

1。2  实验方法 7

1。2。1  植物总RNA的提取 7

1。2。2  逆转录成互补DNA 8

1。2。3  进行PCR扩增出目的片段 9

1。2。4  PCR产物回收 9

1。2。5  提取质粒并进行测序 10

1。2。6  PCR重组载体的构建 10