Expression and purification of N and E proteins from severe acute respiratory syndrome (SARS)-associated coronavirus: a comparative study.
Histidine-tagged N (rNH) and E (rEH) proteins of Severe Acute Respiratory Syndrome (SARS)-coronovirus were expressed in the baculovirus/insect cell system and purified by immobilized metal affinity chromatography. rNH and rEH proteins differed markedly with respect to expression levels, cell death kinetics and subcellular localizations that led to different extraction and purification schemes. The features of both proteins are compared and the potential applications of purified rNH and rEH are discussed.
Lai
Chia-Wei
CW
Department of Chemical Engineering, National Tsing Hua University, 300 Hsinchu, Taiwan.
Chung
Yao-Chi
YC
Lai
Yiu-Kay
YK
Chang
Margaret Dah-Tsyr
MD
Hu
Yu-Chen
YC
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D015153:Blotting, Western; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D002846:Chromatography, Affinity / Q000379:methods; D000086462:Coronavirus Nucleocapsid Proteins; D015870:Gene Expression; D018613:Microscopy, Confocal; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000378:metabolism*; D018411:Spodoptera; D013997:Time Factors; D014162:Transfection; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D000086082:Viroporin Proteins
D000086462:Coronavirus Nucleocapsid Proteins; C501689:E protein, SARS coronavirus; D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D000086082:Viroporin Proteins