Analysis of VSV pseudotype virus infection mediated by rubella virus envelope proteins

Sakata, Masafumi; Tani, Hideki; Anraku, Masaki; Kataoka, Michiyo; Nagata, Noriyo; Seki, Fumio; Tahara, Maino; Otsuki, Noriyuki; Okamoto, Kiyoko; Takeda, Makoto; Mori, Yoshio

experiment. Stock solutions of VSV FLuc -G, VSV FLuc -RV/CE2E1 and RVLP were independently prepared and concentrated and fractionated by sucrose-gradient ultracentrifugation. In this experiment, RVLP, which contains a subgenomic replicon RNA in which the structural genes are replaced with DNA sequences encoding the puromycin N-acetyl-transferase protein, the foot-and-mouth disease virus 2 A self-cleavage domain, and the Renilla luciferase 33 , was used as a marker for RVLP fractionation pattern. VSV FLuc -G, VSV FLuc -RV/CE2E1, and RVLP in individual fractions were detected by infecting Vero cells with the fractions and measuring the relative light unit (RLU) from the cells. The peak infectivity of VSV FLuc -RV/CE2E1 was at ninth fraction similarly to that of VSV FLuc -G, with no peak or shoulder at seventh fraction where RVLP indicated the peak infectivity ( Fig. 2G-I) . The profiles demonstrated that the pseudotype VSV particles, but not RVLP, incorporated the genome.