炭疽杆菌毒力大质粒的驱除及比较蛋白质组学研究

发布时间：2018-10-17 19:54

【摘要】：炭疽芽孢杆菌引发的炭疽病是一种烈性的人畜共患的传染病,危害性极强。尤其近些年,国际上一些恐怖分子利用炭菌杆菌的芽孢制造的恐怖事件,严重干扰了人们的日常生活及社会经济秩序。因此,对其进行研究具有一定的理论价值和潜在的社会效益。在本研究中,运用质粒不相容性原理,分别以炭疽杆菌A16R与A16作为实验的出发菌株,构建了pXO1质粒缺失突变株A16RPI1和pXO2质粒缺失突变株A16PI2。对菌株A16RPI1与A16RP及A16PI2与A16R在生长趋势、糖代谢等方面进行了比较,并鉴定了A16PI2的荚膜生成能力；分别制备了A16RPI1与A16RP的23 h的全菌蛋白质样品和A16PI2与A16R的20 h的全菌蛋白质样品,通过pH 4-7梯度的双向电泳找出差异蛋白,并对差异蛋白点进行了质谱检测分析。得到了以下主要结果： 1)菌株A16RPI1与菌株A16RP的生长趋势基本一致,且二者的糖代谢也无区别。 2)菌株A16PI2与菌株A16R的生长趋势也是一致的,但二者的糖代谢情况稍有不同,A16PI2可以代谢果糖但是A16R却不能代谢果糖。经过荚膜培养实验验证,A16PI2确实不能生成荚膜。 3)菌株A16RPI1与菌株A16RP稳定期蛋白质在pH 4-7这一梯度有5个差异蛋白,这些蛋白都定位在细胞质中,主要是翻译过程中的延伸因子和与氨基酸代谢相关的酶。 4)菌株A16PI2与菌株A16R稳定期蛋白质在pH 4-7这一梯度有22个差异蛋白,这些蛋白基本都定位在细胞质中,只有一个功能未知的蛋白定位在细胞膜上,它们在功能上主要与翻译过程、转录调节、氨基酸的转运和代谢及芽孢的生成相关。
[Abstract]:Anthracnose caused by Bacillus anthracis is a strong zoonotic infectious disease. Especially in recent years, some terrorists in the world have seriously disturbed people's daily life and social economic order by making use of the spores of carbon bacillus. Therefore, its research has certain theoretical value and potential social benefit. In this study, based on the principle of plasmid incompatibility, pXO1 plasmid deletion mutant A16RPI1 and pXO2 plasmid deletion mutant A16PI2 were constructed with Bacillus anthracis A16R and A16 as the starting strains. The growth trend and glucose metabolism of A16RPI1 and A16R were compared with those of A16PI2 and A16R, and the capsule forming ability of A16PI2 was identified, and the whole bacterial protein samples of 23 h of A16RPI1 and A16RP and 20 h of A16PI2 and A16R were prepared, respectively. Differential proteins were identified by pH 4-7 gradient two-dimensional electrophoresis, and the differential protein spots were detected by mass spectrometry. The main results are as follows: 1) the growth trend of strain A16RPI1 and strain A16RP is basically the same, and there is no difference between them. 2) the growth trend of strain A16PI2 and strain A16R is also consistent. However, the glucose metabolism was slightly different between the two groups. A16PI2 could metabolize fructose, but A16R could not metabolize fructose. The results of capsule culture showed that A16PI2 could not produce capsule. 3) there were five differentially expressed proteins in the gradient of stable phase protein of strain A16RPI1 and strain A16RP in the gradient of pH 4-7, all of which were located in the cytoplasm. There are 22 differential proteins between strain A16PI2 and strain A16R stable protein in the gradient of pH 4-7, which are mainly located in cytoplasm. Only one protein with unknown function is located on the cell membrane, which is mainly related to translation, transcriptional regulation, amino acid transport and metabolism, and spore formation.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R378

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