【摘要】：炭疽芽孢桿菌引發(fā)的炭疽病是一種烈性的人畜共患的傳染病,危害性極強(qiáng)。尤其近些年,國際上一些恐怖分子利用炭菌桿菌的芽孢制造的恐怖事件,嚴(yán)重干擾了人們的日常生活及社會(huì)經(jīng)濟(jì)秩序。因此,對(duì)其進(jìn)行研究具有一定的理論價(jià)值和潛在的社會(huì)效益。 在本研究中,運(yùn)用質(zhì)粒不相容性原理,分別以炭疽桿菌A16R與A16作為實(shí)驗(yàn)的出發(fā)菌株,構(gòu)建了pXO1質(zhì)粒缺失突變株A16RPI1和pXO2質(zhì)粒缺失突變株A16PI2。對(duì)菌株A16RPI1與A16RP及A16PI2與A16R在生長(zhǎng)趨勢(shì)、糖代謝等方面進(jìn)行了比較,并鑒定了A16PI2的莢膜生成能力；分別制備了A16RPI1與A16RP的23 h的全菌蛋白質(zhì)樣品和A16PI2與A16R的20 h的全菌蛋白質(zhì)樣品,通過pH 4-7梯度的雙向電泳找出差異蛋白,并對(duì)差異蛋白點(diǎn)進(jìn)行了質(zhì)譜檢測(cè)分析。得到了以下主要結(jié)果： 1)菌株A16RPI1與菌株A16RP的生長(zhǎng)趨勢(shì)基本一致,且二者的糖代謝也無區(qū)別。 2)菌株A16PI2與菌株A16R的生長(zhǎng)趨勢(shì)也是一致的,但二者的糖代謝情況稍有不同,A16PI2可以代謝果糖但是A16R卻不能代謝果糖。經(jīng)過莢膜培養(yǎng)實(shí)驗(yàn)驗(yàn)證,A16PI2確實(shí)不能生成莢膜。 3)菌株A16RPI1與菌株A16RP穩(wěn)定期蛋白質(zhì)在pH 4-7這一梯度有5個(gè)差異蛋白,這些蛋白都定位在細(xì)胞質(zhì)中,主要是翻譯過程中的延伸因子和與氨基酸代謝相關(guān)的酶。 4)菌株A16PI2與菌株A16R穩(wěn)定期蛋白質(zhì)在pH 4-7這一梯度有22個(gè)差異蛋白,這些蛋白基本都定位在細(xì)胞質(zhì)中,只有一個(gè)功能未知的蛋白定位在細(xì)胞膜上,它們?cè)诠δ苌现饕c翻譯過程、轉(zhuǎn)錄調(diào)節(jié)、氨基酸的轉(zhuǎn)運(yùn)和代謝及芽孢的生成相關(guān)。
[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.
【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R378

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