本文選題：芽孢桿菌 + 炭疽�。� 參考：《軍事醫(yī)學》2017年03期

【摘要】：目的構建炭疽芽孢桿菌sigF基因缺失株,以及缺失株的sigF回復株,分析缺失株、回復株表型特點并明確sigF基因對芽孢形成的影響,為后續(xù)研究提供基礎。方法采用同源重組技術在炭疽菌A16D2(pXO1+pXO2~-)菌株sigF基因處插入大觀霉素抗性基因,取代sigF基因,構建了sigF基因缺失突變株。通過構建回復株質粒并電轉到sigF基因缺失株中,構建了sigF基因的回復株。通過生長曲線測定、糖代謝能力比較、顯微鏡觀察芽孢形成及微流控實驗實時觀察細胞生長等方法分析突變株和回復株的特點。結果獲得炭疽菌A16D2菌株sigF基因缺失突變株和相應的回復株。生長曲線測定表明,突變株較出發(fā)菌株在繁殖體階段生長狀態(tài)無明顯差異;糖代謝實驗表明,突變株與出發(fā)菌株在對碳水化合物的利用上差異不顯著;顯微鏡觀察顯示,突變株喪失了形成芽孢的能力、回復株恢復了部分形成芽孢的能力;微流控實驗觀察顯示,缺失株細胞保持在不對稱分裂的狀態(tài),回復株能形成芽孢。結論成功獲得了炭疽菌A16D2sigF基因缺失突變株。該研究證明sigF基因為炭疽菌形成芽孢所必需,但不是細胞生長所必需的因子。
[Abstract]:Objective to construct the sigF gene deletion strain of Bacillus anthracis and the sigF recovery strain of the deletion strain, to analyze the phenotypic characteristics of the deleted strain and to clarify the effect of sigF gene on the spores formation, and to provide a basis for further study. Methods the gene of spectinomycin resistance was inserted into the sigF gene of anthrax strain A16D2(pXO1 pXO _ 2 by homologous recombination technique, replacing the sigF gene, and the mutant of sigF gene deletion was constructed. By constructing the plasmid of the sigF gene and transferring it to the sigF gene deletion strain, the sigF gene recovery strain was constructed. The characteristics of mutants and recovers were analyzed by means of growth curve determination, comparison of glucose metabolism ability, observation of spores formation by microscope and real-time observation of cell growth by microfluidic experiments. Results the sigF gene deletion mutants and corresponding recovers of anthrax A16D2 strain were obtained. The growth curve test showed that there was no significant difference in the growth status of the mutant strain compared with the original strain in the reproductive phase, the glucose metabolism test showed that there was no significant difference in the utilization of carbohydrate between the mutant strain and the original strain, and the microscopic observation showed that there was no significant difference between the two strains in the utilization of carbohydrates. The mutant lost the ability to form spores, and the recovery strain recovered part of the ability to form spores, and the microfluidic experiments showed that the cells of the missing strain remained in the state of asymmetric division, and the recovered strain could form spores. Conclusion the mutant of A16D2sigF gene deletion of anthrax was successfully obtained. This study demonstrated that the sigF group is necessary for the formation of spores by anthrax, but is not an essential factor for cell growth.
【作者單位】： 上海海洋大學食品學院;軍事醫(yī)學科學院生物工程研究所病原微生物生物安全國家重點實驗室;
【基金】：國家自然科學基金資助項目(81271785)
【分類號】：R378.72

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