title: The fusion site of envelope fragments from each serotype of Dengue virus in the P64k protein, influence some parameters of the resulting chimeric constructs.
abstract: 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.
authors.affiliation: División de Vacunas, Centro de Ingeniería Genética y Biotecnologi;a, Habana, Cuba. lisset.hermida@cigb.edu.cu
authors.forename: Aída
authors.initials: A
authors.lastname: Zulueta
authors.forename: Lisset
authors.initials: L
authors.lastname: Hermida
authors.forename: Laura
authors.initials: L
authors.lastname: Lazo
authors.forename: Iris
authors.initials: I
authors.lastname: Valdés
authors.forename: Rayner
authors.initials: R
authors.lastname: Rodríguez
authors.forename: Carlos
authors.initials: C
authors.lastname: López
authors.forename: Ricardo
authors.initials: R
authors.lastname: Silva
authors.forename: Delfina
authors.initials: D
authors.lastname: Rosario
authors.forename: Jorge
authors.initials: J
authors.lastname: Martín
authors.forename: María G
authors.initials: MG
authors.lastname: Guzmán
authors.forename: Gerardo
authors.initials: G
authors.lastname: Guillén
mesh_terms: 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*
chemical_list: D000956:Antigens, Viral; D001425:Bacterial Outer Membrane Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C112623:lpdA protein, Neisseria meningitidis