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biogate21铁虫 (正式写手)
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10个金币求助 High Level Expression of Recombinant Proteins in&n
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以下是High Level Expression of Recombinant Proteins in&n的部分内容 High Level Expression? Protein Biochemistry Westerns and pull-down assays Enzyme Assays Sub-mg quantities Protein Structure Structural Biologists NMR and X-ray crystallography 10’s of mg quantities Part I. Expression Systems Expression System Components Target Protein mRNA cDNA genomic DNA Expression Vector (plasmids) pET pQE pGEX Expression Strain (E. coli ) BL21(DE3) Rosetta(DE3) plus pRARE Tuner(DE3) lacZY deletion mutant ADA494(DE3) trxB mutant Origami(DE3) trxB and gor mutations Expression Plasmid Elements Origin of Replication (ColE1) Selectable marker (ampicillin, kanamycin or chloramphenicol resistance) Multiple cloning site Transcription promoter (T7, T5 or Tac) Regulated expression of target protein Regulation of Protein Expression in the lac Operon pET Expression System Most comprehensive commercial expression system Uses non-native T7 RNA polymerase ~33 different vectors available Regulation of Protein Expression in the pET System Double induction by IPTG T7 RNA polymerase (98 kDa) target gene (only in T7lac vectors) Highly specific Compatible with a wide range of expression hosts requires DE3 lysogen Compatibility with Toxic Genes T7 RNA polymerase inhibited by T7 lysozyme Drawbacks to pET system |
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biogate21
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2楼2009-03-26 14:03:40