Site-specific fluorescence labelling of recombinant polyomavirus-like particles.
For the development of gene therapy protocols based on polyomavirus-like particles, we describe a method for fluorescence labelling of virions in order to study virus-cell interactions preceding gene delivery. Site-specific fluorescence labelling of polyomavirus-like particles is achieved via a single cysteine residue and maleimide conjugates of fluorescence dyes (fluorescein, Texas Red). Polyomavirus-like particles can be assembled in vitro from recombinant capsomers produced in E. coli. Since the assembly process is independent of disulfide bond formation, all cysteine residues of the wild-type protein were replaced by serines, and a new unique cysteine residue was introduced for the attachment of the fluorescence marker.
Schmidt
U
U
Institut für Biotechnologie, Martin-Luther-Universität Halle-Wittenberg, Halle (Saale), Germany.
Kenklies
J
J
Rudolph
R
R
Böhm
G
G
D000818:Animals; D002213:Capsid / Q000737:chemistry; D036022:Capsid Proteins; D002460:Cell Line; D005453:Fluorescence; D005456:Fluorescent Dyes; D051379:Mice; D011120:Polyomavirus / Q000737:chemistry; D014771:Virion; D014966:Xanthenes
D036022:Capsid Proteins; D005456:Fluorescent Dyes; C062481:VP1 protein, polyomavirus; D014966:Xanthenes; C034657:Texas red