[The development and optimization of coupled cell-free expression system for production of the transmembrane domain of the receptor tyrosine kinase ErbB3].
The cell-free expression system based on bacterial extract S30 from E. coli for production of the transmembrane domain of human receptor tyrosine kinase ErbB3 (residues 632-675) was developed. The synthesis of the domain in the soluble form in the presence of detergents and in the form of the translation mixture precipitate was studied. The protocols of purification of the recombinant domain obtained by both methods were developed. The final yield of target protein in optimal conditions was 1.8-2.0 mg per 1 ml of translation mixture.
N F
NF
Khabibullina
E N
EN
Liukmanova
G S
GS
Kopeina
Z O
ZO
Shenkarev
A S
AS
Arsen'ev
D A
DA
Dolgikh
M P
MP
Kirpichnikov
D002474:Cell-Free System; D002846:Chromatography, Affinity; D003902:Detergents / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008567:Membranes, Artificial; D010957:Plasmids; D015202:Protein Engineering / Q000379:methods*; D020893:Receptor, ErbB-3 / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003902:Detergents; D008567:Membranes, Artificial; D011994:Recombinant Proteins; D020893:Receptor, ErbB-3