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摘要：细菌是通过共同合成的AHL信号分子来感知种群密度的。在合作过程中，不可避免产生突变个体，它们不产生信号分子，却利用别的细菌的信号分子。突变个体的产生有利于个体的扩散，却不利于整个种群的生存。细菌是如何在整体水平上抑制突变菌株的产生的，是本课题的研究内容。本课题利用引物扩增pUC18质粒的Apr基因片段，转化后用AatⅡ和EcoR1双酶切，得到两端粘性末端长为866的片段；用酶切技术酶切pPop-galac质粒，一份用Ava1和AatⅡ双酶切，得到3526的大片段回收；另一份用EcoR1和Ava1双酶切，得到556bp小片段回收。混合三者连接得到新构建的工程菌模型，但是在酶切556bp片段时还存在诸多问题。24554
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毕业论文关键词：合作行为、群体感应、群体密度、阈值
A construction of cooperative plasmid based on Quorum-sensing
Abstract: The cooperation behavior between organisms have important influence on its evolution. The engineering bacterium model constructed by this topic will start the related gene expression in the population density reaches a threshold, resulting in double resistance. The bacteria perceive the population density through the synthesis of AHL signal molecules. Inevitable in the process of cooperation mutated inpiduals, they don't produce signaling molecules, but the use of other bacteria signal molecule . Mutation conducive to the generation of inpidual dispersal, but not conducive to the survival of the species. How bacteria inhibition mutant strain on the overall level is the research content of this topic. Apr gene fragment was amplified by pUC18 primers of this subject, with AatⅡ and EcoR1 double enzyme digestion after transformation, get two sticky end length of 866 fragments. By restriction enzyme digestion of plasmid pPop-galac technology, a Ava1 and AatⅡ double enzyme digestion, high recovery 3526; another with EcoR1 and Ava1 double enzyme digestion, 556bp small fragment recovery. Mixed three and connected the new-building engineering strain , but there are still many problems in the digested 556bp fragments.
Key words: cooperation behavior, quorum sensing, population density, threshold value
目录
毕业设计(论文) 1
1. 前言 1
1.1细菌的群体感应现象 1
1.2革兰氏阴性细菌的群体感应系统 2
1.3本课题的研究思路 2
1.4本课题的主要内容和研究意义 3
2. 实验材料与方法 4
2.1材料与设备仪器 4
2.1.1实验材料 4
2.1.2主要的仪器设备 5
2.1.3实验所用菌种 5
2.1.4培养基和试剂的制备 6
2.2实验方法 7
2.2.1 菌种的扩大培养 7
2.2.2 提取质粒 7
2.2.3 PCR扩增 8
2.2.4 T-载体连接 9
2.2.5目的质粒的转化 9
2.2.6质粒的酶切 9
2.2.7 琼脂糖凝胶电泳 11
2.2.8 琼脂糖凝胶的回收 11
3 实验方案、结果与分析 12
3.1实验方案 13
3.2实验结果与分析 14
3.2.1提取质粒 14
3.2.2 PCR扩增 15
3.2.3 T-载体连接 16
3.2.4目的质粒的转化及验证 17
3.2.5质粒的酶切 17
3.2.6目的条带的割胶回收 21
4. 实验结论 23
致谢 24
参考文献 25
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