Imaging of vaccinia virus entry into HeLa cells.
The recently developed technique of live cell imaging has found numerous applications, including the detection of virus movements in living cells. To monitor virus motility, viruses or cellular proteins are fused with fluorescence markers and then detected by time-lapse fluorescence microscopy. These techniques allow kinetic analyses of individual virus particles in motion during the virus entry process as well as monitoring of dynamic interactions between viruses and cellular structures in real time. The methods presented here describe how to construct a fluorescent recombinant vaccinia virus expressing the core protein A4L fused to mCherry, and how to detect the virus movement on actin-EYFP-expressed HeLa cells.
Academia Sinica, Institute of Molecular Biology, Taipei, Taiwan, ROC.
Cheng-Yen
CY
Huang
Wen
W
Chang
D000199:Actins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D018929:Cell Culture Techniques; D003001:Cloning, Molecular; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D059010:Single-Cell Analysis / Q000379:methods; D059008:Time-Lapse Imaging / Q000379:methods*; D014616:Vaccinia virus / Q000254:growth & development / Q000302:isolation & purification / Q000502:physiology*; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism; D014776:Virus Cultivation; D053586:Virus Internalization; D000097573:Red Fluorescent Protein
D000199:Actins; D001426:Bacterial Proteins; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D014758:Viral Core Proteins; D000097573:Red Fluorescent Protein; C054562:yellow fluorescent protein, Bacteria