In vitro assembly of nucleocapsid-like particles from purified recombinant capsid protein of dengue-2 virus.
The capsid protein is one of the three structural proteins of flaviviruses and is the building block of the nucleocapsid. It has also a predominant role in the replication of dengue virus. To obtain nucleocapsid-like particles from recombinant dengue-2 capsid protein produced in E. coli, a purification process using cation exchange chromatography was established. The purified protein exhibited a molecular mass corresponding to a dimer; therefore, similar to that reported for alphaviruses, an in vitro assembly reaction using single-stranded DNA was performed. In all cases, particles were obtained independently of the specificity and the length of the oligonucleotides used. The present work is the first report of in vitro assembly of the recombinant dengue capsid protein, which could constitute a powerful tool in the development of vaccine candidates.
López
Carlos
C
Center for Genetic Engineering and Biotechnology, Playa, Havana, Cuba.
Gil
Lázaro
L
Lazo
Laura
L
Menéndez
Ivón
I
Marcos
Ernesto
E
Sánchez
Jorge
J
Valdés
Iris
I
Falcón
Viviana
V
de la Rosa
María C
MC
Márquez
Gabriel
G
Guillén
Gerardo
G
Hermida
Lisset
L
D036022:Capsid Proteins / Q000235:genetics / Q000378:metabolism*; D004277:DNA, Single-Stranded / Q000378:metabolism; D003716:Dengue Virus / Q000502:physiology*; D004926:Escherichia coli / Q000235:genetics; D046529:Microscopy, Electron, Transmission; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D022701:Virosomes / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D019065:Virus Assembly
D036022:Capsid Proteins; D004277:DNA, Single-Stranded; D011994:Recombinant Proteins; D022701:Virosomes