Blocking the dengue virus 2 infections on BHK-21 cells with purified recombinant dengue virus 2 E protein expressed in Escherichia coli.
Dengue viruses (DVs) are mosquito-borne infectious pathogens. They have become an expanding public health problem in the tropics and subtropics. The dengue envelope (E) protein is one of the viral structure proteins responsible mainly for the virus attachment and entry onto host cells. It is also the major immunogen for virus neutralization. In this study, we have constructed a recombinant plasmid expressing a truncated E protein of DV-2 virus PL046 strain. The C-terminal hydrophobic domain of the E protein was removed and replaced with the sequence of S peptide to facilitate expression and purification. When expressed in Escherichia coli, the recombinant E proteins were found to be in the form of aggregated state. Through denaturation and dialysis processes, the receptor-interacting function of the purified recombinant E proteins was maintained, which was demonstrated by its ability to inhibit the DV-2 plaque-forming efficiency on mammalian BHK-21 host cells.
Chiu
Mei-Wui
MW
Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan, ROC.
Yang
Yun-Liang
YL
D000818:Animals; D001667:Binding, Competitive; D002460:Cell Line; D006224:Cricetinae; D003716:Dengue Virus / Q000254:growth & development / Q000472:pathogenicity*; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D012995:Solubility; D014759:Viral Envelope Proteins / Q000235:genetics*; D010948:Viral Plaque Assay
C051821:E-glycoprotein, Dengue virus type 2; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins