Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S Gerstein opyright◎2001 ISBNS:0-471-37972-7( Paper);0-47 (Electronic) E coli Expression Systems Peter a. bell Expression Vector Structure What Makes a Plasmid an Expression Vector? Is a Stronger Promoter Always Desirable Why Do Promoters Leak and What Can You Do What Factors Affect the level of translation? 464 What Can Affect the Stability of the Protei the cell? Which Protein Expression System Suits Your Needs? Track Record What Do You Know about the Gene to Be Expressed?..465 Very Promising. What Levels of Expression snould o..468 What Do You know about Your protein? Advertisements for Commercial Expression Vectors Ar :r60 47 Why Should You Select a Fusion System? 471 When Should You Avoid a Fusion System? Is It Necessary to Cleave the Tag off the Fusion Protein? 474 Will Extra Amino acid Residues Affect Your protein of Interest after Digestion? Working with Expression Systems 475 What Are the Options for Cloning a Gene for Expression?... 475461 15 E. coli Expression Systems Peter A. Bell Expression Vector Structure . . . . . . . . . . . . . . . . . . . . . . . . . . . . 462 What Makes a Plasmid an Expression Vector? . . . . . . . . . . . 462 Is a Stronger Promoter Always Desirable? . . . . . . . . . . . . . . 463 Why Do Promoters Leak and What Can You Do about It? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 464 What Factors Affect the Level of Translation? . . . . . . . . . . . . 464 What Can Affect the Stability of the Protein in the Cell? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 464 Which Protein Expression System Suits Your Needs? . . . . . . . 465 Track Record . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 465 What Do You Know about the Gene to Be Expressed? . . . 465 What Do You Know about Your Protein? . . . . . . . . . . . . . . . 468 Advertisements for Commercial Expression Vectors Are Very Promising. What Levels of Expression Should You Expect? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 470 Which E. coli Strain Will Provide Maximal Expression for Your Clone? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 471 Why Should You Select a Fusion System? . . . . . . . . . . . . . . . 471 When Should You Avoid a Fusion System? . . . . . . . . . . . . . . . 472 Is It Necessary to Cleave the Tag off the Fusion Protein? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 474 Will Extra Amino Acid Residues Affect Your Protein of Interest after Digestion? . . . . . . . . . . . . . . . . . . . . . . . . . . . . 475 Working with Expression Systems . . . . . . . . . . . . . . . . . . . . . . . 475 What Are the Options for Cloning a Gene for Expression? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 475 Molecular Biology Problem Solver: A Laboratory Guide. Edited by Alan S. Gerstein Copyright © 2001 by Wiley-Liss, Inc. ISBNs: 0-471-37972-7 (Paper); 0-471-22390-5 (Electronic)