High-efficiency expression and purification of the soluble glycoprotein extracellular domain of Rabies virus strain CTN / 中华微生物学和免疫学杂志

Objective To express and purify the glycoprotein extracellular domain (Ex-GP) of Rabies virus strain CTN in soluble form with high efficiency.Methods A recombinant expression plasmid containing the gene encoding the Ex-GP was constructed.Various expression conditions were screened to obtain an optimum prokaryotic expression system for Ex-GP in soluble form.The expressed target protein was purified using affinity chromatography and gel filtration chromatography.Results The target protein Ex-GP with high antigenicity was efficiently expressed in soluble form by using the recombinant PBCX expression system and effectively purified by using affinity and gel filtration chromatography.Conclusion The soluble form of Ex-GP is successfully expressed and purified in a simple and convenient way.This study paves the way for further researches on the biological functions of rabies virus glycoprotein,the pathogenic mechanism of rabies and the development of diagnostic reagent and vaccines for rabies virus.

Expression and purification of the soluble envelope protein of Japanese encephalitis virus / 中华微生物学和免疫学杂志

Objective To express and purify the envelope ( E) protein of Japanese encephalitis virus (JEV) in soluble form.Methods Various prokaryotic expression vectors , host strains and induction conditions including time and temperatures were screened to obtain an optimum prokaryotic expression system for JEV E protein in soluble form .The expressed protein was purified by using nickel column chromatography and gel filtration chromatography .Results The soluble JEV E protein , accounting for 23% of the totally bacteria soluble protein was effectively expressed by using the recombinant plasmid PBCX -E406 at low tem-perature.The purity of the expressed protein reached up to 85%after the purification by using nickel column and gel filtration chromatography .Conclusion Soluble JEV E protein was successfully expressed and puri-fied in a simple and efficient way .It would provide a useful tool for further investigation on JEV infection , attenuation mechanism of JE live vaccine strain SA 14-14-2 and the quality control of JE vaccine .It can also be used for the development of diagnosis assay for JEV .