Translation Initiation: Extract Systems and Molecular by Jon Lorsch

By Jon Lorsch

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Gualerzi, unpublished observations). , 2001). For the same reason, the use of His-tagged factors is not recommended. All three initiation factor genes were engineered to make their expression more reliable and efficient. , 1993) and infC (Brombach and Pon, 1987) genes were separated from the neighboring genes in their transcriptional units that repress initiation factor expression. The translationally autorepressed AUU triplet of infC was substituted with the canonical AUG (Brombach and Pon, 1987).

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Dipeptide Formation 9. Applications of the Method 10. Binding of fMet-tRNA to the 30S Subunit 11. 1016/S0076-6879(07)30001-3 # 2007 Elsevier Inc. All rights reserved. 1 2 Pohl Milon et al. 12. Nucleotide Binding to IF2 13. Subunit Joining Acknowledgments References 25 25 26 26 Abstract Initiation of mRNA translation in prokaryotes requires the small ribosomal subunit (30S), initiator fMet-tRNAfMet, three initiation factors, IF1, IF2, and IF3, and the large ribosomal subunit (50S). During initiation, the 30S subunit, in a complex with IF3, binds mRNA, IF1, IF2·GTP, and fMet-tRNAfMet to form a 30S initiation complex which then recruits the 50S subunit to yield a 70S initiation complex, while the initiation factors are released.

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