1. 毕业设计（论文）的内容和要求

论文内容 本研究以大肠杆菌为研究对象，采用分子克隆技术，构建糖基转移酶ugt73g1与蔗糖合酶sus共表达重组质粒，采用乳糖诱导培养基诱导表达产酶，利用全细胞催化手段催化槲皮素合成槲皮素-3，4#8217;-二葡萄糖糖苷。

同时，在全细胞催化反应中，研究各条件因素对双酶偶联反应体系中底物转化率的影响，以及对目标产物进行定量分析。

论文要求 （1）掌握文献查阅的一般方法，学会在中国期刊网、web of science科学引文索引、springer link电子期刊、elsevier sdos电子期刊等检索资源上查阅关于生物催化槲皮素研究的相关文献，并对大肠杆菌表达系统有全面了解。

2. 参考文献

[1] Son, Mi Hye , Bong-Gyu Kim , Dae Hwan Kim . Production of Flavonoid O-Glucoside Using Sucrose Synthase and Flavonoid O-Glucosyltransferase Fusion Protein. J. Microbiol. Biotechnol. (2009), 19(7), 709#8211;712. [2] Yi Li, Sandie Baldauf, Eng-Kiat Lim, and Dianna J. Bowles. Phylogenetic Analysis of the UDP-glycosyltransferase Multigene Family of Arabidopsis thaliana. THE JOURNAL OF BIOLOGICAL CHEMISTRY. (2001), Vol. 276, No. 6, 4338#8211;4343. [3] Joe Ross, Yi Li, Eng-Kiat Lim and Dianna J Bowles. Higher plant glycosyltransferases. Genome Biology. 2001, 2(2): reviews3004.1#8211;3004.6. [4] Catherine M. Kramera , Rogerio T.N. Prataa , Michael G. Willitsa , Vincenzo De Lucab , John C. Steensa , Gerson Grasera, *. Cloning and regiospecificity studies of two flavonoid glucosyltransferases from Allium cepa. Phytochemistry. 64 (2003) 1069#8211;1076. [5] Masao Hirotani, Ryo Kuroda, Hideyuki Suzuki, Takafumi Yoshikawa. Cloning and expression of UDP-glucose: avonoid 7-O-glucosyltransferase from hairy root cultures of Scutellaria baicalensis. Planta (2000) 210: 1006-1013. [6] Frederik De Bruyn, Jo Maertens, Joeri Beauprez, Wim Soetaert, Marjan De Mey. Biotechnological advances in UDP-sugar based glycosylation of small molecules. Biotechnology Advances. 33 (2015) 288#8211;302. [7] Lavoie EG, Kukulski F, Levesque SA, Lecka J, Sevigny J. 2004. Cloning and characterization of mouse nucleoside triphosphate diphosphohydrolase-3. Biochem Pharmacol. 67:1917#8211;1926. [8] Eng-Kiat Lim, David A. Ashford, Bingkai Hou, Rosamond G. Jackson, Dianna J. Bowles. Arabidopsis Glycosyltransferases as Biocatalysts in Fermentation for Regioselective Synthesis of Diverse Quercetin Glucosides. Garfield Weston Foundation for CNAP. 2004. [9] Pesnot T, Jorgensen R, Palcic MM, Wagner GK. 2010. Structural and mechanistic basis for a new mode of glycosyltransferase inhibition. Nat Chem Biol. 6:321#8211;323. [10] Teng Y, Liu Q, Ma J, Liu F, Han Z, Wang Y, Wang W. 2006. Cloning, expression and characterization of a novel human CAP10-like gene hCLP46 from CD34( ) stem/progenitor cells. Gene. 371:7#8211;15.

3. 毕业设计（论文）进程安排

2018.2.26-2018.3.2 做预实验，给后期实验提供指导意义； 2018.3.3-2018.3.9 查阅文献，并结合预实验结果分析，设计合理实验方案； 2018.3.10-2018.5.4 将已有的含有双酶的重组质粒导入到大肠杆菌中，采用乳糖诱导培养基诱导产酶；初步建立双酶催化槲皮素合成槲皮素-3，4#8217;-二葡萄糖糖苷体系； 2018.5.5-2018.5.31 研究各条件因素对全细胞反应体系底物转化率的影响，并合理优化提高产物产率，同时及时整理、分析实验数据； 2018.6.1-2018.6.15 及时撰写毕业论文。