A convenient method for genetic incorporation of multiple noncanonical amino acids into one protein in Escherichia coli.
By overexpressing the C-terminal domain of the ribosomal protein L11 to decrease release factor 1-mediated termination of protein translation, enhanced amber suppression is achieved in E. coli. This enhanced amber suppression efficiency allows the genetic incorporation of three N(epsilon)-acetyl-l-lysines into one GFP(UV) protein in E. coli.
Huang
Ying
Y
Department of Chemistry, Texas A&M University, College Station, TX, 77843, USA.
Russell
William K
WK
Wan
Wei
W
Pai
Pei-Jing
PJ
Russell
David H
DH
Liu
Wenshe
W
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D018388:Codon, Terminator; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D005798:Genes, Bacterial; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008239:Lysine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012269:Ribosomal Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D049490:Systems Biology
D000596:Amino Acids; D018388:Codon, Terminator; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D012269:Ribosomal Proteins; C027624:ribosomal protein L11; D049452:Green Fluorescent Proteins; C016949:N-epsilon-acetyllysine; D008239:Lysine