Protein Synthesis

1. Front Matter

   Pages i-xiv

   PDF

2. Analyses of Ribosome Distribution During In Vitro Translation

   • Sandra L. Wolin

   Pages 1-9

3. Allele-Specific Priming in the Mapping of rRNA

   • Stephen Douthwaite

   Pages 11-21

4. Permeabilized Mammalian Cells as a System for Protein Synthesis

   • Romualdas Stapulionis, Murray P. Deutscher

   Pages 23-31

5. The Measurement of Processivity Errors in Protein Synthesis

   • Hengjiang Dong

   Pages 33-42

6. A Biosynthetic Approach for the Incorporation of Unnatural Amino Acids into Proteins

   • Jon S. Thorson, Virginia W. Cornish, Jeannie E. Barrett, Sharon T. Cload, Takato Yano, Peter G. Schultz

   Pages 43-73

7. The Analysis of Translational Activity Using a Reporter Gene Constructed from Repeats of an Antibody-Binding Domain from Protein A

   • Asgeir Björnsson, Salim Mottagui-Tabar, Leif A. Isaksson

   Pages 75-91

8. In Vitro Engineering Using Acyl-Derivatized tRNAs

   • Obed W. Odom, Wieslaw Kudlicki, Boyd Hardesty

   Pages 93-103

9. In Vitro Engineering Using Synthetic tRNAs with Altered Anticodons Including Four-Nucleotide Anticodons

   • Gisela Kramer, Wieslaw Kudlicki, Boyd Hardesty

   Pages 105-116

10. Analysis of rRNA Function Using Specialized Ribosomes

    • Marcel F. Brink, Herman A. de Boer

    Pages 117-128

11. Determination of the Peptide Elongation Rate In Vivo

    • Michael A. Sørensen, Steen Pedersen

    Pages 129-142

12. The Measurement of Missense Errors During In Vivo Protein Synthesis

    • Bernard N. Violand, Gregg Bogosian

    Pages 143-151

13. Heelprinting Analysis of In Vivo Ribosome Pause Sites

    • Margaret J. Hollingsworth, Jeong-Kook Kim, Neil E. Stollar

    Pages 153-165

14. Analysis of elF-2α Kinases in Yeast

    • Thomas E. Dever

    Pages 167-178

15. Continuous-Flow Cell-Free Translation, Transcription-Translation, and Replication-Translation Systems

    • Lyubov A. Ryabova, lgor Yu. Morozov, Alexander S. Spirin

    Pages 179-193

16. Analysis of Translational Activity of Extracts Derived from Oocytes and Eggs of Xenopus laevis

    • Virginia M. Pain, Tina D. Patrick, Richard Cox, Simon J. Morley

    Pages 195-209

17. A Fractionated Reticulocyte Lysate System for Studies on Protein Synthesis Initiation Factors

    • Michael Rau, Theophile Ohlmann, Virginia M. Pain, Simon J. Morley

    Pages 211-226

18. Measurement of Ribosomal Accuracy and Proofreading in E. coli Burst Systems

    • Måns Ehrenberg, Neş’e Bilgin

    Pages 227-241

19. Measurement of Rate of Protein Synthesis In Vitro

    • Neş’e Bilgin, Måns Ehrenberg

    Pages 243-256

20. In Vivo Mutational Analysis of Ribosomal RNA in Saccharomyces cerevisiae

    • Jaap Venema, Rudi J. Planta, Hendrik A. Raué

    Pages 257-270

The synthesis of proteins from 20 or so constituent amino acids according to a strictly defined code with an accuracy of better than 1 in 10,000 at most loca­ tions is arguably the most complex task performed by cells. Protein Synthesis collects together methods and protocols covering a range of different approaches towards understanding how the cellular machinery accomplishes this task and how these ftinctions might be harnessed by the biotechnology industry to generate novel and useful proteins. The era in which the components of the translational machinery were being catalogued is over. This volume gathers together protocols that focus on preserving and describing the dynamic function as closely as possible. The need to understand exactly how ribosomes are positioned on messages or where tRNA molecules, translation factors, or control proteins are bound, has been appreciated by many of the authors. Several chapters that explore the fidelity and processivity of translation reflect this belief. Moreover, the fundamental importance of rRNA at the heart of the ribosome is a strong theme in a number of the protocols. These articles include in vitro and in vivo systems from bacterial, fungal, plant, and animal systems. Overall, Protein Synthesis might be characterized by the novelty of the approaches employed to illuminate the inner workings of the protein synthetic machinery as well as by the inventiveness of the attempts to harness these reactions for biotechnological applications.

". . . an excellent protocols book, a copy of which should be on the shelves of every molecular biology lab."-Microbiology Today

• Blond McIndoe Center, Queen Victoria Hospital, Sussex, England

  Robin Martin

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