title: High-level expression of tamavidin 2 in human cells by codon-usage optimization.
abstract: Tamavidin 2 is a fungal protein that binds to biotin with an extremely high affinity. Tamavidin 2 is superior to avidin or streptavidin in terms of its low-level non-specific binding and high-level thermal stability. However, the gene for tamavidin 2 is highly expressed in Escherichia coli but not in mammalian cells, restricting its application as an affinity tag in mammalian cells. Here, we optimized the codon usage of tamavidin 2 for human cells and found that the resultant mutant expressed tamavidin 2 at approximately 30-fold higher level compared with the native gene. The protein thus produced in human cells could be purified by iminobiotin affinity chromatography, bound tightly to biotin, and was stable at high temperature (82 °C). This powerful technology for high-level expression of tamavidin 2 in mammalian cells will be of value in evaluating various fusion proteins produced in mammalian cells for numerous applications.
authors.affiliation: a Plant Innovation Center , Japan Tobacco Inc. , 700 Higashibara, Iwata, Shizuoka 438-0802 , Japan.
authors.forename: Yoshimitsu
authors.initials: Y
authors.lastname: Takakura
authors.forename: Sakurako
authors.initials: S
authors.lastname: Katayama
authors.forename: Yuki
authors.initials: Y
authors.lastname: Nagata
mesh_terms: D000595:Amino Acid Sequence; D001360:Avidin / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001710:Biotin / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002846:Chromatography, Affinity; D003062:Codon / Q000737:chemistry / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D005818:Genetic Engineering; D057809:HEK293 Cells; D006358:Hot Temperature; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D055550:Protein Stability; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D003062:Codon; D005656:Fungal Proteins; D011994:Recombinant Proteins; C550582:tamavidin 2, Pleurotus cornucopiae; D001360:Avidin; D001710:Biotin
keywords: biotin; codon usage; expression; mammalian cell; tamavidin