Soluble protein expression in E. coli cells using IgG-binding domain of protein A as a solubilizing partner in the cold induced system.
We constructed a cold induced expression vector in Escherichia coli cells that consists of a histidine tag sequence for nickel chelate affinity purification, IgG-binding domain of protein A (ZZ-domain) and the multiple cloning sites. The role of ZZ-domain as a solubilizing partner at 15 degrees C was demonstrated by expressing the imidazopyrazinone-type luciferases of Renilla, Oplophorus, Gaussia, and Vargula (Cypridina) as well as the calcium-binding photoproteins and firefly luciferase. The fused protein with ZZ-domain was expressed efficiently as a soluble form in the cytoplasm of E. coli cells at low temperature.
Inouye
Satoshi
S
Yokohama Research Center, Chisso Corporation, 5-1 Okawa, Kanazawa-ku, Yokohama 236-8605, Japan. sinouye@chisso.co.jp
Sahara
Yuiko
Y
D000331:Aequorin / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001059:Apoproteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D022762:CCAAT-Enhancer-Binding Proteins / Q000235:genetics; D003080:Cold Temperature; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006360:Heat-Shock Proteins / Q000235:genetics; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D008156:Luciferases / Q000235:genetics; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012995:Solubility; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D001059:Apoproteins; D022762:CCAAT-Enhancer-Binding Proteins; D006360:Heat-Shock Proteins; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; C094867:YBX3 protein, human; C060014:apoaequorin; D000331:Aequorin; D008156:Luciferases