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1. 毕业设计（论文）的内容和要求
1.文献阅读与归纳，深入了解课题背景2.谷氨酸棒杆菌敲除质粒的构建3.掌握生物膜表征的方法和发酵操作4.独立完成毕业论文的撰写
2. 实验内容和要求
1 掌握谷氨酸棒杆菌基因敲除和表达的方法2 掌握发酵实验及产物检测的基本操作3 掌握用结晶紫测量生物量的方法
3. 参考文献
1. Jiang Y, Qian F, Yang J, et al. CRISPR-Cpf1 assisted genome editing of Corynebacterium glutamicum[J]. Nature communications, 2017, 8: 15179.2. Zhang J, Yang F, Yang Y, et al. Optimizing a CRISPR-Cpf1-based genome engineering system for Corynebacterium glutamicum[J]. Microbial cell factories, 2019, 18(1): 60.3. Zetsche B, Gootenberg J S, Abudayyeh O O, et al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system[J]. Cell, 2015, 163(3): 759-771.4. Flemming H C, Wingender J, Szewzyk U, et al. Biofilms: an emergent form of bacterial life[J]. Nature Reviews Microbiology, 2016, 14(9): 563.5. Cho J S, Choi K R, Prabowo C P S, et al. CRISPR/Cas9-coupled recombineering for metabolic engineering of Corynebacterium glutamicum[J]. Metabolic engineering, 2017, 42: 157-167.6. Chen C, Li Y, Hu J, et al. Metabolic engineering of Corynebacterium glutamicum ATCC13869 for L-valine production[J]. Metabolic engineering, 2015, 29: 66-75.7. Boles B R, Horswill A R. Agr-mediated dispersal of Staphylococcus aureus biofilms[J]. PLoS pathogens, 2008, 4(4): e1000052.8. Podder M S, Majumder C B. Predictive approach for simultaneous biosorption and bioaccumulation of arsenic by Corynebacterium glutamicum MTCC 2745 biofilm supported on NL/MnFe2O4 composite[J]. Journal of Water Process Engineering, 2016, 11: 8-31.9. Klompe S E, Vo P L H, Halpin-Healy T S, et al. Transposon-encoded CRISPRCas systems direct RNA-guided DNA integration[J]. Nature, 2019, 571(7764): 219-225.10. Wang T, Li Y, Li J, et al. An update of the suicide plasmid‐mediated genome editing system in Corynebacterium glutamicum[J]. Microbial biotechnology, 2019.11. Muffler A, Bettermann S, Haushalter M, et al. Genome-wide transcription profiling of Corynebacterium glutamicum after heat shock and during growth on acetate and glucose[J]. Journal of biotechnology, 2002, 98(2-3): 255-268.12. Engels S, Schweitzer J E, Ludwig C, et al. clpC and clpP1P2 gene expression in Corynebacterium glutamicum is controlled by a regulatory network involving the transcriptional regulators ClgR and HspR as well as the ECF sigma factor σH[J]. Molecular microbiology, 2004, 52(1): 285-302.
4. 毕业设计（论文）计划
2020.12-2021.01 下载并阅读与课题相关的文献，完成开题报告2021.02-2021.03 利用CRISPR技术构建谷氨酸棒杆菌突变菌株2021.03-2021.04突变菌株生物膜表征和固定化发酵实验2021.04-2021.06 整理实验数据并撰写毕业论文
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