Journal: bioRxiv
Article Title: Transgene Expression Kinetics and Replication Potential of Recombinant Adenovirus Serotype 4 in a Mouse Model and its Use as a Herpes Simplex Virus Vaccine
doi: 10.64898/2026.05.15.725395
Figure Lengend Snippet: Construction and characterization of Ad4 expressing luciferase or HSV-2 gD. (A) The E3 region of the adenovirus serotype 4 (Ad4) genome is replaced by left and right arms (LA, RA) to allow recombination with a cassette with LA, a CMV promoter, a luciferase or HSV-2 gD gene, a bovine growth hormone (BGH) poly A site, and an RA. The recombinant Ad4 genome is approximately 37 kb in length and retains genes in the E1 region responsible for viral replication. The genome map abbreviations are as follows; left inverted terminal repeat (LITR), E1-early transcriptional gene region 1 (consisting of E1A and E1B), E2-early transcriptional gene region 2 (consisting of E2A and E2B), E3-early transcriptional gene region 3, E4-early transcriptional gene region 4, and right inverted terminal repeat (RITR). (B) In vitro Ad4-Luc infection of A549 cells showing cytopathic effects (MOI 0.1, 3 days after infection, 5x magnification). (C) Uninfected A549 cells for comparison (5x magnification). (D) Relative luminescence of A549 cells infected with Ad4-Luc, Ad4-EGFP, or uninfected. Bars show median values with interquartile range (IQR), with individual data points. ## p=0.009 (Ad4-Luc versus Ad4-EGFP); ** p=0.002 (Ad4-Luc versus uninfected); ns: not significant. (E) Western blot of purified soluble HSV-2 gD protein (Sol gD2), cell lysates of A549 cells infected with Ad4-gD2 (Ad4-gD2), a replicate lysate (Ad4-gD2 rep), and uninfected A549 cells after staining with anti-HSV-2 gD antibody.
Article Snippet: The previously described SpeI site was utilized to linearize pRAd4-EGFP or pRAd4-Luc and this was transfected using PEIpro® (Polyplus Cat# 101000033) into the A549 human carcinoma lung cell line (ATCC cat. CCL-185).
Techniques: Expressing, Luciferase, Recombinant, In Vitro, Infection, Comparison, Western Blot, Purification, Staining