1. 毕业设计（论文）的内容和要求
论文内容为提高乳清酸到三磷酸胞苷( CTP) 的转化效率, 克隆并表达了编码 CTP合成酶( CTPS) 的基因pyrG。
从大肠杆菌44102中克隆出三磷酸胞苷合成酶基因（pyrG）克隆到pet28a载体上，并转入大肠杆菌E.coli Rosetta (DE3) 中表达，优化了诱导，温度，PH等条件，测定该酶的酶学性质，实现三磷酸胞苷合成酶的高效表达。
论文要求1．文献查阅掌握文献查阅，使用网络资源如中国期刊网、维普数据库、超星数字图书馆、Elsevier电子期刊、Springer Link全文电子期刊等检索资源，查阅关于包涵体蛋白复性的相关知识。
2. 参考文献
1. Huang, M. and L.M. Graves, De novo synthesis of pyrimidine nucleotides; emerging interfaces with signal transduction pathways. Cell Mol Life Sci, 2003. 60(2): p. 321-36.2. Endrizzi, J.A., et al., Crystal structure of Escherichia coli cytidine triphosphate synthetase, a nucleotide-regulated glutamine amidotransferase/ATP-dependent amidoligase fusion protein and homologue of anticancer and antiparasitic drug targets. Biochemistry, 2004. 43(21): p. 6447-63.3. Iyengar, A. and S.L. Bearne, Aspartate-107 and leucine-109 facilitate efficient coupling of glutamine hydrolysis to CTP synthesis by Escherichia coli CTP synthase. Biochem J, 2003. 369(Pt 3): p. 497-507.4. Fijolek, A., A. Hofer, and L. Thelander, Expression, purification, characterization, and in vivo targeting of trypanosome CTP synthetase for treatment of African sleeping sickness. Journal of Biological Chemistry, 2007. 282(16): p. 11858-11865.5. Lunn, F.A., J.E. MacDonnell, and S.L. Bearne, Structural requirements for the activation of Escherichia coli CTP synthase by the allosteric effector GTP are stringent, but requirements for inhibition are lax. Journal of Biological Chemistry, 2008. 283(4): p. 2010-2020.6. Scheit, K.H. and H.-J. Linke, Substrate Specificity of CTP Synthetase from Escherichia coli. European Journal of Biochemistry, 1982. 126(1): p. 57-60.7. Anderson, P.M., CTP synthetase from Escherichia coli: an improved purification procedure and characterization of hysteretic and enzyme concentration effects on kinetic properties. Biochemistry, 1983. 22(13): p. 3285-3292.8. Ostrander, D.B., et al., Effect of CTP synthetase regulation by CTP on phospholipid synthesis in Saccharomyces cerevisiae. Journal of Biological Chemistry, 1998. 273(30): p. 18992-18992.9. Park, T.-S., et al., Identification of Ser424 as the protein kinase A phosphorylation site in CTP synthetase from Saccharomyces cerevisiae. Biochemistry, 1999. 38(27): p. 8839-8848.10. Meng, Q. and R.L. Switzer, Regulation of Transcription of the Bacillus subtilis pyrG Gene, Encoding Cytidine Triphosphate Synthetase. Journal of Bacteriology, 2001. 183(19): p. 5513-5522.11. Jrgensen, C.M., et al., Expression of the pyrG gene determines the pool sizes of CTP and dCTP in Lactococcus lactis. European Journal of Biochemistry, 2004. 271(12): p. 2438-2445.12. Han, G.-S., et al., Expression of human CTP synthetase in Saccharomyces cerevisiae reveals phosphorylation by protein kinase A. Journal of Biological Chemistry, 2005. 280(46): p. 38328-38336.13. Goto, M., et al., Crystal structures of CTP synthetase reveal ATP, UTP, and glutamine binding sites. Structure, 2004. 12(8): p. 1413-23.14. Lauritsen, I., et al., Structure of the dimeric form of CTP synthase from Sulfolobus solfataricus. Acta Crystallographica Section F, 2011. 67(2): p. 201-208.15. Lunn, F.A., T.J. MacLeod, and S.L. Bearne, Mutational analysis of conserved glycine residues 142, 143 and 146 reveals Gly142 is critical for tetramerization of CTP synthase from Escherichia coli. Biochemical Journal, 2008. 412(1): p. 113-121.16. Willemoes, M. and B.W. Sigurskjold, Steady-state kinetics of the glutaminase reaction of CTP synthase from Lactococcus lactis. The role of the allosteric activator GTP incoupling between glutamine hydrolysis and CTP synthesis. Eur J Biochem, 2002. 269(19): p. 4772-9.17. MacLeod, T.J., F.A. Lunn, and S.L. Bearne, The role of lysine residues 297 and 306 in nucleoside triphosphate regulation of E. coli CTP synthase: Inactivation by 2′,3′-dialdehyde ATP and mutational analyses. Biochimica et Biophysica Acta (BBA) - Proteins 2): p. 248-254.21. Lineweaver, H. and D. Burk, The Determination of Enzyme Dissociation Constants. Journal of the American Chemical Society, 1934. 56(3): p. 658-666.22. 丁晓慧, 花放, 刘文瑞等. CGRP对大鼠缺血再灌注后海马内突触体素表达的影响. 解剖科学进展. 2000, 6(4): 359~360.23. Jahn R, Sdhof T C. Synaptic vesicle traffic：rush hour in the never terminal.J. Neurochem. 1993, 61: 12~2124. Alberghina M. Lipid peroxidation inhibits acyl-CoA：-1-a-cyi-sn-hiycero-3-Phosphocholine O-acyltransferase but not CTP：phosphocholine cytidylyltransferase activity in rat brain membranes. Neurochem. Int.1995, 26: 477~487.25.李纳新, 董敬远, 张建华. 三磷酸胞苷二钠与毕奥星治疗急性脑血管病的疗效观察. 河南使用神经疾病杂志. 2003,7,6(4): 76~77.26 李骏, 梁富元, 何锋. 降纤酶、美络宁联合治疗急性脑梗塞50例. 右江民族医学院学报. 1999, 6: 923~924.27. 徐叔云主编.现代实用临床药理学 [M].北京：华夏出版社.1996: 546~54828.陈信成, 冯刚民, 杨晓荣. 胞二磷胆碱与三磷酸胞苷二钠治疗急性脑梗死120例疗效比较. 右江医学. 2003, 31(1)29. 王进敏, 沈文省. 三磷酸胞苷二钠治疗脑出血的观察与护理. 齐鲁护理杂志. 2003，9(10)30. 周爱儒等主编.生物化学.北京：人民卫生出版社，2002: 123~130.31. 邓常青等. 补阳还五汤及其有效部位对沙鼠脑缺血后DND脑能量代谢、NO和NOS的影响. 中药药理与临床. 1999, 15: 5.32. 曹华政, 李贤彬, 熊震等. Sichuan Medical Journal，2003，Vol.24，No.333. 冯永歌, 刘建军等. 三磷酸胞苷二钠治疗新生儿缺氧缺血性脑病34例. 河南医科大学学报. 2001, 36(4): 494~495.34. 钟乃川. 突发性耳聋与高海拔红细胞增多症.中华耳鼻咽喉科志.1980, 15: 58.35 尹述成，李俊，金康业. 突发性聋预后的相关因素浅析. 听力学及言语疾病杂志. 1998, 6: 202.36. Maier S F, Wiertlak E P, Watkins L R. Endogenous pain facilitory system. Am Pain Soc J. 1992, 1:191~198.
以上是毕业论文任务书，课题毕业论文、开题报告、外文翻译、程序设计、图纸设计等资料可联系客服协助查找。
