Choice of Cellular Protein Expression System

Abstract

Recombinant protein expression has become a standard laboratory tool, and a wide variety of systems and techniques are now in use. Because there are so many systems to choose from, the investigator has to be careful to use the combination that will give the best results for the protein being studied. This overview unit discusses expression and production choices, including post-translational modifications (e.g., glycosylation, acylation, sulfation, and removal of N-terminal methionine), in vivo and in vitro folding, and influence of downstream elements on expression.

Ebola virus VP35 induces high-level production of recombinant TPL-2–ABIN-2–NF-κB1 p105 complex in co-transfected HEK-293 cells

“…However, protein yields are usually much lower than in most commonly used non-mammalian expression hosts, such as Escherichia coli , yeast and baculovirus-infected insect cells [29]. …”

“…However, achievable yields in HEK-293 cells are far lower than possible with E. coli and baculovirus expression systems [29]. The activation of PKR by transfected DNA plasmids decreases translation and limits recombinant protein production by HEK-293 cells [40].…”

Expressão, purificação e caracterização parcial de proteínas relacionadas à patogenicidade de Magnaporthe grisea

“…Visto que a concentração de sal, a temperatura, a aeração do meio de cultura regulada pela velocidade de rotação e o pH do tampão de extração, bem como agentes caotrópicos como 4M uréia podem influir de modo considerável na solubilidade de proteínas, diferentes testes de solubilização foram realizados com as proteínas XYL5 e XYL5.DOM (Gray et al, 2000;Mayer, 2004;Tresaugues et al, 2004) . Com o uso de uréia, pôde-se observar a solubilização, mas não o refolding correto das proteínas,…”