Design of carrier tRNAs and selection of four-base codons for efficient incorporation of various nonnatural amino acids into proteins in Spodoptera frugiperda 21 (Sf21) insect cell-free translation system

This article describes the chemical aminoacylation of the yeast phenylalanine suppressor tRNA with a series of amino acids bearing fluorinated side chains via the hybrid dinucleotide pdCpA and ligation to the corresponding truncated tRNA species. Aminoacyl-tRNAs can be used to synthesize biologically relevant proteins which contain fluorinated amino acids at specific sites by means of a cell-free translation system. Such engineered proteins are expected to contribute to our understanding of discrete fluorines’ interaction with canonical amino acids in a native protein environment and to enable the design of fluorinated proteins with arbitrary desired properties.

Download Full-text

Incorporation of Unnatural Non-α-Amino Acids into the N Terminus of Proteins in a Cell-Free Translation System

ChemBioChem ◽

10.1002/cbic.200700249 ◽

2007 ◽

Vol 8 (14) ◽

pp. 1650-1653 ◽

Cited By ~ 18

Author(s):

Norihito Muranaka ◽

Masanori Miura ◽

Hikaru Taira ◽

Takahiro Hohsaka

Keyword(s):

Amino Acids ◽

Translation System ◽

N Terminus ◽

Free Translation ◽

A Cell

Download Full-text

Incorporation of fluorescently labeled nonnatural amino acids into proteins in an E. coli in vitro translation system

Nucleic Acids Symposium Series ◽

10.1093/nass/3.1.271 ◽

2003 ◽

Vol 3 (1) ◽

pp. 271-272 ◽

Cited By ~ 8

Author(s):

T. Hohsaka ◽

R. Abe ◽

K. Shiraga ◽

M. Sisido

Keyword(s):

Amino Acids ◽

In Vitro Translation ◽

Translation System ◽

Nonnatural Amino Acids ◽

E Coli

Download Full-text

Position-specific incorporation of biotinylated non-natural amino acids into a protein in a cell-free translation system

Biochemical and Biophysical Research Communications ◽

10.1016/j.bbrc.2007.07.099 ◽

2007 ◽

Vol 361 (3) ◽

pp. 794-799 ◽

Cited By ~ 17

Author(s):

Takayoshi Watanabe ◽

Norihito Muranaka ◽

Issei Iijima ◽

Takahiro Hohsaka

Keyword(s):

Amino Acids ◽

Translation System ◽

Free Translation ◽

A Cell ◽

Natural Amino Acids

Download Full-text

A Concept for Selection of Codon-Suppressor tRNAs Based on Read-Through Ribosome Display in anIn VitroCompartmentalized Cell-Free Translation System

Journal of Nucleic Acids ◽

10.1155/2012/538129 ◽

2012 ◽

Vol 2012 ◽

pp. 1-7 ◽

Cited By ~ 3

Author(s):

Atsushi Ogawa ◽

Masayoshi Hayami ◽

Shinsuke Sando ◽

Yasuhiro Aoyama

Keyword(s):

Stop Codon ◽

Full Length ◽

Translation System ◽

Anticodon Loop ◽

Ribosome Display ◽

Amber Stop Codon ◽

Free Translation ◽

Translatable Mrna ◽

Selection Of

Here is presented a concept forin vitroselection of suppressor tRNAs. It uses a pool of dsDNA templates in compartmentalized water-in-oil micelles. The template contains a transcription/translation trigger, an amber stop codon, and another transcription trigger for the anticodon- or anticodon loop-randomized gene for tRNASer. Upon transcription are generated two types of RNAs, a tRNA and a translatable mRNA (mRNA-tRNA). When the tRNA suppresses the stop codon (UAG) of the mRNA, the full-length protein obtained upon translation remains attached to the mRNA (read-through ribosome display) that contains the sequence of the tRNA. In this way, the active suppressor tRNAs can be selected (amplified) and their sequences read out. The enriched anticodon (CUA) was complementary to the UAG stop codon and the enriched anticodon-loop was the same as that in the natural tRNASer.

Download Full-text