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毕业论文网 > 任务书 > 化学化工与生命科学类 > 生物工程 > 正文

来自赤红球菌的P450单加氧酶的克隆及表达研究任务书

 2020-06-24 19:51:12  

1. 毕业设计(论文)的内容和要求

在赤红球菌(rhodococcus ruber )dsm44319菌株中,一个iv型细胞色素p450单加氧酶家族的新成员被识别出来。

在赤红球菌的基因组还没有被测序时,一个类似p450基因的片段就已经用简并引物扩增出来。

类似p450的dna片段的侧翼区域,则使用聚合酶链式反应,以基因组步行的方式识别出来。

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2. 参考文献

[1] Altschul SF.Amino acid substitution matrices from an information theoretic perspective [J].J Mol Biol,1991,219:555-565. [2] Bornscheuer UT. Methods to increase enantioselectivity of lipases and esterases [J]. Curr Opin Biotechnol,2002,13:543-547. [3] Bornscheuer UT, Pohl M. Improved biocatalysts by directed evolution and rational protein design[J].Curr Opin Chem Biol,2001,5:137#8211;143. [4] Cryle MJ, Stok JE, De Voss JJ.Reactions catalyzed by bacterial cytochromes P450[J].Aust J Chem,2003,56:749-762. [5] De Mot R, Parret AH.A novel class of self-sufficient cytochrome P450 monooxygenases in prokaryotes[J].Trends Microbiol,2002,10:502-508. [6] Farinas ET, Bulter T, Arnold FH.Directed enzyme evolution[J].Curr Opin Biotechnol,2001,12:545-551. [7] Finnerty WR.The biology and genetics of the genus Rhodococcus[J]. Annu Rev Microbiol,1992,46:193-218. [8] Fruetel JA, Mackman RL, Peterson JA, Ortiz de Montellano PR.Relationship of active site topology to substrate specificity for cytochrome P450terp (CYP108)[J]. J Biol Chem,1994,269:28815-28821. [9] Gliszczynska A,Koziolowa A. Chromatographic determination of flavin derivatives in baker#8217;s yeast[J].J Chromatogr A,1998,822:59-66. [10] Gustafsson MC, Roitel O,et al.Expression, purification, and characterization of Bacillus subtilis cytochromes P450 CYP102A2 and CYP102A3: flavocytochrome homologues of P450 BM3 from Bacillus megaterium[J].Biochemistry,2004,43:5474-5487. [11] Harford-Cross CF, Carmichael AB, Allan FK, England PA, Rouch DA, Wong LL.Protein engineering of cytochrome P450 (cam) (CYP101) for the oxidation of polycyclic aromatic hydrocarbons[J].Protein Eng,2010,13:121-128. [12] Hedegaard J, Gunsalus IC.Mixed function oxidation. IV. An induced methylene hydroxylase in camphor oxidation.[J].J Biol Chem ,1965,240:4038-4043. [13] Li QS, Ogawa J, Schmid RD, Shimizu S.Engineering cytochrome P450 BM-3 for oxidation of polycyclic aromatic hydrocarbons[J].Appl Environ Microbiol,2001, 67:5735-5739. [14] Turner NJ, Flitsch SL.A self-sufficient cytochrome P450 with a primary structural organization that includes a flavin domain and a [2Fe#8211;2S] redox center[J].J Biol Chem,2003,278:48914-48920. [15] Schmid A, Dordick JS, Hauer B, Kiener A, Wubbolts M, Witholt B. Industrial biocatalysis today and tomorrow[J].Nature,2001,409:258#8211;268. [16] Seng Wong T, Arnold FH, Schwaneberg U. Laboratory evolution of cytochrome P450 BM-3 monooxygenase for organic cosolvents[J].Biotechnol Bioeng,2004,85:351-358. [17] Werck-Reichhart D, Feyereisen R. Cytochromes P450: a success story[J].Genome Biol,2000,1:REVIEWS3003.

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 2018.1.1-1.10 选题,与导师讨论题目 2018.1.11 导师下达任务书 2018.1.12-2018.2.8 根据任务书要求,查找相关文献资料,完成开题报告 2018.02.9-05.23 完成试验并提交论文第一稿 2018.05.24-05.27 根据导师意见修改论文第一稿,完成并提交第二稿 2018.05.28-06.1 根据导师意见修改论文第二稿,完成并提交第三稿 2018.06.1-06.7 完成论文的审阅,准备答辩

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