1. 毕业设计（论文）的内容和要求
内容：对插入4HBD基因的重组丙酮丁醇梭菌菌株进行培养及发酵实验，验证能否产一定量的1,4-丁二醇。
同时进行基因敲除实验，检验相关的基因敲除对产量的影响。
要求：掌握微生物实验的基本规范操作和遵守实验室的安全规章制度
2. 参考文献
1.Van Dien S: From the first drop to the first truckload:commercialization of microbial processes for renewablechemicals. Curr Opin Biotechnol 2013, 24:1061-1068.2. Yim H, Haselbeck R, Niu W, Pujol-Baxley C, Burgard A, Boldt J, Khandurina J, Trawick JD, Osterhout RE, Stephen R et al.:Metabolic engineering of Escherichia coli for directproduction of 1,4-butanediol. Nat Chem Biol 2011, 7:445-452.3.Dhamankar H, Prather KL: Microbial chemical factories: recentadvances in pathway engineering for synthesis of value added chemicals. Curr Opin Struct Biol 2011, 21:488-494.4.Trinh CT: Elucidating and reprogramming Escherichia coli metabolisms for obligate anaerobic n-butanol and isobutanol production. Appl Microbiol Biotechnol 2012, 95:1083-1094.4. Navarrete C, Nielsen J, Siewers V: Enhanced ethanol production and reduced glycerol formation in fps1 mutants of Saccharomyces cerevisiae engineered for improved redox balancing. AMB Express 2014, 4:86.5. Liu J, Qi H, Wang C, Wen J: Model-driven intracellular redox status modulation for increasing isobutanol production in Escherichia coli. Biotechnol Biofuels 2015, 8:108.6.Bettenbrock K, Bai H, Ederer M, Green J, Hellingwerf KJ, Holcombe M, Kunz S, Rolfe MD, Sanguinetti G, Sawodny O et al.:Towards a systems level understanding of the oxygenresponse of Escherichia coli. Adv Microb Physiol 2014,64:65-114.7. Yang TH: 13C-based metabolic flux analysis: fundamentals and practice. Methods Mol Biol 2013, 985:297-334.8.Barton NR, Burgard AP, Burk MJ, Crater JS, Osterhout RE,Pharkya P, Steer BA, Sun J, Trawick JD, Van Dien SJ et al.: An integrated biotechnology platform for developing sustainable chemical processes. J Ind Microbiol Biotechnol 2015, 42:349-360.9. Patnaik R: Engineering Complex Phenotypes in Industrial Strains. John Wiley 2013.10.Fletcher E, Krivoruchko A, Nielsen J: Industrial systems biology and its impact on synthetic biology of yeast cell factories. Biotechnol Bioeng 2015, 113:1164-1170. 11.Tai YS, Xiong M, Jambunathan P, Wang J, Wang J, Stapleton C Zhang K: Engineering nonphosphorylative metabolism to generate lignocellulose-derived products. Nat Chem Biol 2016, 12:247-253.12. Clark DP: The fermentation pathways of Escherichia coli. FEMS Microbiol Rev 1989, 5:223-234.13. Kurihara S, Kato K, Asada K, Kumagai H, Suzuki H: A putrescineinducible pathway comprising PuuE-YneI in which gammaaminobutyrate is degraded into succinate in Escherichia coli K-12. J Bacteriol 2010, 192:4582-4591.14. Pham VD, Somasundaram S, Lee SH, Park SJ, Hong SH: Engineering the intracellular metabolism of Escherichia coli to produce gamma-aminobutyric acid by co-localization of GABA shunt enzymes. Biotechnol Lett 2015, 38:321-327.15. Sousa PM, Silva ST, Hood BL, Charro N, Carita JN, Vaz F, Penque D, Conrads TP, Melo AM: Supramolecular organizations in the aerobic respiratory chain of Escherichia coli. Biochimie 2011, 93:418-425.16. Sousa PM, Videira MA, Bohn A, Hood BL, Conrads TP, Goulao LF, Melo AM: The aerobic respiratory chain of Escherichia coli: from genes to supercomplexes. Microbiology 2012, 158:2408-2418.17. Benoit S,AbaibouH,Mandrand-Berthelot MA: Topological analysis of the aerobic membrane-bound formate dehydrogenase of Escherichia coli. J Bacteriol 1998, 180:6625-6634.
