The fusion site of envelope fragments from each serotype of Dengue virus in the P64k protein, influence some parameters of the resulting chimeric constructs.
To characterize the effect of the envelope fragment fusion site in the P64k protein from Neisseria meningitidis several chimeric constructs were obtained. One variant consisted in the insertion of the E fragment from each Dengue serotype within the lipoil binding domain of the P64k, whereas the other was based on the fusion of the envelope fragment at the C-terminus of the same meningoccocal protein. The results of the expression study revealed the majoritary levels with the C-terminus fusion variants of each serotype. In contrast, the highest proportion of soluble protein was reached with the insertion variants independently of the viral serotype. On the other hand, a significant level of degradation was detected for the semipurified forms of the insertion variants being remarkable in the Dengue 2 construct. Finally, the recognition by Dengue murine antibodies was similar independently of the fusion site. Regarding these results, we can affirm the suitability of the C-terminus fusion variants for further vaccine development as well as for a diagnostic system.
Zulueta
Aída
A
División de Vacunas, Centro de Ingeniería Genética y Biotecnologi;a, Habana, Cuba. lisset.hermida@cigb.edu.cu
Hermida
Lisset
L
Lazo
Laura
L
Valdés
Iris
I
Rodríguez
Rayner
R
López
Carlos
C
Silva
Ricardo
R
Rosario
Delfina
D
Martín
Jorge
J
Guzmán
María G
MG
Guillén
Gerardo
G
D000956:Antigens, Viral / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D003716:Dengue Virus / Q000145:classification / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology*
D000956:Antigens, Viral; D001425:Bacterial Outer Membrane Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C112623:lpdA protein, Neisseria meningitidis