Codon optimization of the BirA enzyme gene leads to higher expression and an improved efficiency of biotinylation of target proteins in mammalian cells.
Biotinylation of proteins is an attractive alternative to 'epitope-tagging', due to the strong biotin-(strept)avidin interaction and to the wide commercial availability of reagents for detection and purification of biotinylated macromolecules. Enzymatic biotinylation of target proteins in vivo using short biotin acceptor domains was described previously. Their use in mammalian cell requires expression of the bacterial biotinylation enzyme BirA. Here we describe the construction of a humanized version of BirA, with most of the rare codons replaced by codons that are more frequently used in human cells. The humanized BirA is expressed better in mammalian cells, resulting in improved efficiency of biotinylation in vivo. We anticipate that the humanized BirA gene will find use in many applications that involve in vivo biotinylation.
Mechold
Undine
U
CNRS UPR 9079, Laboratoire Oncogenèse, Différenciation et Transduction du Signal, Institut André Lwoff, 7 rue Guy Moquet, Bat B, Villejuif 94800, France.
Gilbert
Cristele
C
Ogryzko
Vasily
V
D019921:Biotinylation / Q000379:methods*; D019731:Carbon-Nitrogen Ligases / Q000096:biosynthesis* / Q000235:genetics*; D002460:Cell Line; D003062:Codon / Q000235:genetics; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D024861:Genetic Enhancement / Q000379:methods; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014157:Transcription Factors / Q000096:biosynthesis* / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D003062:Codon; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D019731:Carbon-Nitrogen Ligases; C058125:birA protein, E coli