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Fig. 1 | Virology Journal

Fig. 1

From: Optimization of Zika virus envelope protein production for ELISA and correlation of antibody titers with virus neutralization in Mexican patients from an arbovirus endemic region

Fig. 1

Design and production of Zika Envelope proteins. a. Design of ZIKV envelope constructs. The full genome of ZIKV is shown. The genes for pre-membrane (prM) and envelope (Env) including the 3 domains DI, DII, DIII, stem (S) and transmembrane regions (TM) are highlighted. The red arrow indicates the direction of transcription. 8 plasmid constructs encoding prM-Env or Env were designed and cloned into either pOPINTTGneo or pOPINTTGneo-3C-CD4 expression vectors resulting in total of 16 constructs. b. The structure of secreted Env proteins: (top) prM-Env (1–589) in pOPINTTGneo produces Env protein with a His-tag after cleavage of prM upon secretion. (bottom) prM-Env (1–589) in pOPINTTGneo-3C-CD4 produces Env protein with 3C protease site, CD4 fusion tag and His tag. The approximate sizes of components of Env and Env-CD4 proteins are shown in kDa. c. An example of western-blot of Env-CD4 (62.5 kDa) and Env (44 kDa) secreted from HEK293T and detected with anti-His antibodies for constructs encoding prM-Env and prM-Env-CD4 are shown. The positive control was the ectodomain of cell surface receptor sFcεRIα [46] and the negative control was GFP. d. Coomassie Brilliant Blue staining (CBB) of the fractions of purified Env-CD4 and Env proteins are shown

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