1. 毕业设计（论文）的内容和要求

论文内容 人肿瘤坏死因子α(tnf-α)是一种涉及全身炎症和免疫反应的多功能细胞因子。

它由各种人细胞特别是活化的巨噬细胞和单核细胞产生。

其最显著的活性特征是可以在体内或体外特异地杀伤多种肿瘤细胞，对正常细胞则无明显的细胞毒作用。

2. 参考文献

1. Horiuchi T , Mitoma H , Harashima S , et al. Transmembrane TNF-α: structure, function and interaction with anti-TNF agents[J]. Rheumatology, 2010, 49(7):1215-1228. 2. Ma D , Chen Y , Fang L , et al. Purification and characterization of RGD tumor-homing peptide conjugated human tumor necrosis factor α over-expressed in Escherichia coli[J]. Journal of Chromatography B Analytical Technologies in the Biomedical Life Sciences, 2007, 857(2):231-239. 3. Osaki T , Nakanishi T , Aoki M , et al. Soluble Expression in Escherichia coli of a Single-Domain Antibody#8211;Tumor Necrosis Factor α Fusion Protein Specific for Epidermal Growth Factor Receptor[J]. Monoclonal Antibodies in Immunodiagnosis Immunotherapy, 2018, 37(1):20. 4. Papaneophytou C P , Kontopidis G A . Optimization of TNF-α overexpression in Escherichia coli using response surface methodology: Purification of the protein and oligomerization studies[J]. Protein Expression and Purification, 2012, 86(1):35-44. 5. Tsukamoto H , Fukudome K , Kohara J , et al. Preparation of recombinant murine tumor necrosis factor-alpha in Escherichia coli: a rapid method to remove tags from fusion proteins by thrombin-cleavage and ion-exchange chromatography.[J]. Protein Expression Purification, 2007, 56(1):138-144. 6. Liu Y , Cheung L H , Marks J W , et al. Recombinant single-chain antibody fusion construct targeting human melanoma cells and containing tumor necrosis factor[J]. International Journal of Cancer, 2004, 108(4):549-557. 7. Halin C , Gafner V , Villani M E , et al. Synergistic therapeutic effects of a tumor targeting antibody fragment, fused to interleukin 12 and to tumor necrosis factor alpha[J]. Cancer Research, 2003, 63(12):3202-3210. 8. Wang Y , Ren W , Gao D , et al. One-step refolding and purification of recombinant human tumor necrosis factor-α (rhTNF-α) using ion-exchange chromatography.[J]. Biomedical Chromatography, 2015, 29(2):305-311. 9. Cheng C , Wu S , Cui L , et al. A novel Ffu fusion system for secretory expression of heterologous proteins in Escherichia coli[J]. Microbial Cell Factories, 2017, 16(1):231. 10. 吴珊珊，朱芸，陈珊珊，何冰芳. 融合标签在蛋白质可溶性表达中的应用进展[J]. 化工进展, 2014, 33(4):993-998.

3. 毕业设计（论文）进程安排

起讫日期 设计（论文）各阶段工作内容 备 注 2018.12.26-2019.1.18 文献查阅，完成开题报告 2019.3.1-2019.3.30 引物设计，基因扩增 2019.4.1-2019.4.30 构建重组TNF-α表达质粒 2019.5.5-2019.5.28 重组TNF-α表达条件优化 2019.5.39-2019.6.6 毕业论文撰写、答辩