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摘要：生物之间的合作行为对其进化规律有着重要的影响。基于群体感应的细菌行为具有明显的社会性。本课题构建的工程菌模型，会在种群密度达到阈值时启动相关基因表达，从而利用乳糖碳源，实现二次生长。本课题论文通过将这种群体感应质粒转化大肠杆菌后，在不同条件下观察其生长规律及其原因，为建立合作种群模型提供依据。实验结果显示：在好氧、TBK环境中，加IPTG，乳糖组最先达到稳定期，且稳定期OD600值远小于只加乳糖组，空白对照组生长最具优势；在厌氧、TBK环境下，延长时间至2860MIN，大肠杆菌还未达到稳定期，且加IPTG、乳糖生长最差，只加乳糖组生长最好；在厌氧、M9环境下，IPTG、葡萄糖组与葡萄糖组最先达到稳定期，IPTG、葡萄糖、乳糖组与葡萄糖、乳糖组同时到稳定期；转化一个月后，工程菌在含乳糖、X-gal的LB平板上变蓝，而刚转化的工程菌在含乳糖、X-gal的LB平板上却是白色菌落。因此，这样构建的质粒对于通过IPTG实现群体感应还存在诸多问题。5542
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关键词：合作行为、群体感应、群体密度、阈值、二次生长
Effect verification of gene cycle: imitated quorum sensing cooperative behavior
Abstract: The cooperation behavior between biology has an important influence on its evolution law. Bacterial behavior has obvious sociality base on quorum sensing. The model of engineering bacteria constructed in this paper can active related gene and use lactose to realize secondary growth when the population density reach threshold value. This paper will discuss engineering bacteria after transformed quorum sensing plasmid growth regulation and reason under different environment and provide evidence for the establishment of cooperation population model. The result depicted: the IPTG, lactose supplemented reached stable phase first and its bacteria density was lowest under the environment of TBK, oxygen; In the environment of anaerobic and TBK, the IPTG, lactose supplemented grow worst, lactose only grow best, but no one reach stable phase even we added grow time to 2860MIN; The IPTG, glucose and glucose supplemented reach stable phase first in the anaerobic M9, the IPTG, lactose, glucose and lactose, glucose supplemented reach stable phase at the same time; After one month from transform, the engineering bacteria showed blue colonies in LB plate supplemented lactose, X-gal, but bacteria showed white colonies one month before. Therefore, there are problems for IPTG inducing constructed plasmid and realizing quorum sensing.
Keyword: cooperation behavior, quorum sensing, population density, threshold value, secondary growth
目录
毕业设计(论文) 1
1. 前言 1
1.1微生物合作关系 1
1.2微生物的利他行为 1
1.3微生物的限制性扩散与群体感应现象 1
1.4基于群体感应的细菌利他合作行为 2
1.5本课题的研究思路与主要内容 2
2. 实验材料与方法 3
2.1 材料与设备仪器 3
2.11实验材料 4
2.12主要的仪器设备 5
2.13实验所用菌种 5
2.14培养基和试剂的制备 5
2.2实验方法 6
2.21菌种的扩大培养 6
2.22提取质粒 6
2.23琼脂糖凝胶电泳 7
2.24质粒的酶切 8
2.25琼脂糖凝胶的回收 8
2.26酶切产物的连接 9
2.27目的质粒的转化 10
2.28目的质粒的验证 10
2.29工程菌株的生长曲线 11