《第8章基因表达教学材料》由会员分享，可在线阅读，更多相关《第8章基因表达教学材料（31页珍藏版）》请在金锄头文库上搜索。
1、第八章 克隆基因的表达Expression of Cloned GenesGene Organization单击此处编辑母版标题样式单击此处编辑母版副标题样式*3Gene Expression and Regulation LevelsStructure and expression of the mammalian -globin geneTranscription and TranslationTranscription in Prokaryotes Structure of a typical promoter in E.coli开始转录开始转录T T G A C AT T G A C
2、AA A C T G TA A C T G T-35 -35 区区(Pribnow box)(Pribnow box)T A T A A T Pu T A T A A T Pu A T A T T A PyA T A T T A Py-10 -10 区区1 1-30-30-50-501010-10-10-40-40-20-205 5 3 3 3 3 5 5 原核生物启动子保守序列原核生物启动子保守序列RNA-polRNA-pol识别位点识别位点(recognition site) (recognition site) 5 5 5 5 RNARNA聚合酶保护区聚合酶保护区结构基因结构基因3 3
3、3 3 RNA转录起始-35区-10区TTGACATTAACTTTTACATATGATTTTACATATGTTTTGATATATAATCTGACGTACTGTN17N16N17N16N16N7N7N6N7N6AAAAAtrp tRNATyrlacrecAAra BAD TTGACA TTGACA TATAAT TATAAT共有序列 TATA TATA盒盒 CAAT CAAT盒盒 GC GC盒盒 增强子增强子 顺式作用元件顺式作用元件 结构基因结构基因-GCGC-CAAT-TATA-GCGC-CAAT-TATA转录起始转录起始真核生物启动子保守序列真核生物启动子保守序列RNARNA聚合酶全酶在转
4、录起始区的结合聚合酶全酶在转录起始区的结合 P Ptactac启动子启动子 P Ptrptrp与与 P Placlac的杂化产物的杂化产物 P Ptactac比原来各自的活性强比原来各自的活性强 -35-35区区 -10 -10区区 Ptrp TTGACA TTAACT Plac TTTACA TATGTT 共有序列共有序列 TTGACA TATAATTTGACA TATAAT P Ptac17tac17 TTGACA TATAATTTGACA TATAAT S-D Recognition Sequences in Prokaryotic mRNAsTermination of RNA Cha
5、insExpression VectorSecretion Expression VectorFusional ExpressionVector Factor Xa-Activated form of factor x that participates in both the intrinsic and extrinsic pathways of blood coagulation. It catalyses the conversion of prothrombin to thrombin in conjunction with other cofactors. Conserved Seq
6、uences in PromotersDistance of Structure Genes and PromotersGene expression level and secondarystructure of mRNAAlteration of the Distance between Gene and PromoterTranslation Reading FrameConstruction the Reading FrameInduced Expression of Gene Chemical-lac promoter and IPTG IPTG即Isopropyl -D-1-thi
7、ogalactopyranoside，也称Isopropyl -D-thiogalactoside，中文名为异丙基 -D-硫代半乳糖苷。分子式为C9H18O5S ， 分子量为238.30。 -IPTG, cell growth+IPTG, target gene expressionInduced Expression of Gene Physical-PL and temperature sensitive(cI875 lysogen,32,42)Induced Expression of GenePromoter: PLPR Terminator: rrnBT1T2Supression: 32Gene expression: 42 Transcription and Translation in SV40 VirusSV40 Virus VectorSV40 Virus Vector and COS CellsExpression Vectors for Mammalian CellsGene Expressionin Mammals
