摘要绿色光合细菌特有的捕光天线称为绿小体，其被膜上存在着若干小分子量的蛋白质如CsmA，CsmM，CsmN和CsmP。这些蛋白质的结构和功能以及在绿小体的光能传递过程中的作用方式和机理尚待阐明。本研究从克隆关键的基因入手，利用分子生物学和蛋白质化学方法来主要研究其中的CsmP，为研究该基因异源表达的蛋白质提供可靠依据。88257

本实验运用生物信息学分析方法，从已完成测序的JGI全基因组中找出目的基因序列，对蛋白的基本性质进行软件分析，设计特异性引物对其进行PCR扩增，采用新方法进行双标签质粒的构建，之后再转化正确的重组质粒到高效载体Trans1-T1中，进行质粒的扩增，通过菌液检测以及对检测结果正确的菌液进行测序，获得测序结果正确的克隆体，从而完成目标基因的高效克隆，为后续实验与研究提供坚实可靠的基础。

毕业论文关键词：光合细菌；绿小体； CsmP； 基因克隆； 双标签质粒

Abstract Green photosynthetic bacteria contain a novel type of light-harvesting antenna called chlorosome。 In the envelope of chlorosome, there are several kinds of small molecular weight proteins such as CsmA, CsmM, CsmN, and CsmP。 The structure and function of these special proteins and its role in the energy transfer process and its mechanism are yet to be clarified。 In this study, we used the methods of molecular biology and protein chemistry to study the CsmP gene in order to study of the gene and its heterologous expression product to provide reliable basis of the following research work。

This study includes the following experiments: Firstly, by using bioinformatics analysis method, from the JGI whole genome sequence, the target gene sequence has been found out。 Then, the basic properties of translated target protein were analyzed by several online software; And, the specific primers for PCR amplification was devised, especially we used new design of double label in order to construct a plasmid, and then transform the correct recombinant plasmid into high-efficiency carrier Trans1-T1 for amplification。 After identifying by colony PCR and sequencing, the correctly positive recombinant plasmid was obtained。 The high efficiency clone strain was made successfully to meet the target gene cloning, provide reliable basis for follow-up study and research。

Keyword: photosynthetic bacteria; chlorosome; CsmP; gene cloning; double label plasmid

目    录

第一章 引言 5

1。1 研究背景和研究现状 5

1。1。1 研究背景 5

1。1。2 国内外研究现状和发展趋势 5

1。2 研究意义和技术From~优E尔L论E文W网wWw.YoUeRw.com 加QQ7520.18766路线 7

1。2。1 研究的目的及意义 7

1。2。2 实验设计思路 7

1。2。3 技术路线 8

第二章 材料与方法 9

2。1 实验材料 9

2。1。1 主要实验仪器 9

2。1。2 主要实验试剂 9

2。1。3 培养基和实验试剂的配制 10

2。1。4 载体结构图 11

2。2 实验方法 11

2。2。1 设计引物 11

2。2。2 稀释引物 12

2。2。3 PCR扩增CsmP基因以及sGFP基因片段