Technical Reference #1838

1838.

Herpes Simplex Virus 1 Regulatory Protein ICP27 Undergoes a Head-to-Tail Intramolecular Interaction Felicia P. Hernandez and Rozanne M Sandri-Goldin, University of California Irvine School of Medicine, Journal of Virology, 84(1838), (2010)
Link To Paper

Materials & Methods:
BiFC. BiFC analysis was performed in transient transfection assays. The BiFC fusion constructs N-Venus/ICP27/C-Venus N-Venus/ICP27 ICP27/C-Venus ICP27/N-Venus N-Venus/ICP27 (L8A L11A L15A)/C-Venus N-Venus/ ICP27(C483 488S)/C-Venus and N-Venus/ICP27 (S16A S18A)/C-Venus were transfected into RSF cells grown in glass-bottom culture dishes (MatTek Corporation) using Lipofectamine 2000 by following the manufacturer’s protocol. Eighteen hours after transfection cells were infected at a multiplicity of infection (MOI) of 10 with 27-LacZ. Cells were viewed directly for fluorescence using a Zeiss LSM 510 Meta confocal microscope at a magnification of 633.

Microscopic Technique
Confocal Microscopy

Cell Type(s)
Rabbit skin fibroblasts (RSF) and Vero cells