Fig. 1From: Optimization of Zika virus envelope protein production for ELISA and correlation of antibody titers with virus neutralization in Mexican patients from an arbovirus endemic regionDesign 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 shownBack to article page