Author: Choi, Kang-Seuk
Title: Newcastle disease virus vectored vaccines as bivalent or antigen delivery vaccines Document date: 2017_7_26
ID: vk59ghjm_7
Snippet: In general, reverse genetics approaches used to rescue rNDV are based on one of three different systems: (1) a chicken fibroblast cell (or QM5 cell line) and fowl pox virus-T7 recombinant system [13] ; (2) a T7 RNA polymerase expressed BSR T7/5 (baby hamster kidney-21 cell clone) system [14] ; or (3) a human epidermoid carcinoma (HEp-2) cell and a host-restricted recombinant vaccinia virus (modified vaccinia virus Ankara [MVA]) system [15] . Fig......
Document: In general, reverse genetics approaches used to rescue rNDV are based on one of three different systems: (1) a chicken fibroblast cell (or QM5 cell line) and fowl pox virus-T7 recombinant system [13] ; (2) a T7 RNA polymerase expressed BSR T7/5 (baby hamster kidney-21 cell clone) system [14] ; or (3) a human epidermoid carcinoma (HEp-2) cell and a host-restricted recombinant vaccinia virus (modified vaccinia virus Ankara [MVA]) system [15] . Fig. 1 depicts the reverse genetics system used in our own laboratory. Briefly, HEp-2 cells are infected with MVA and then cotransfected with a NDV cDNA clone (LaSota backbone) and three helper plasmids (N, P, and L). The recovered viruses are purified on HEp-2 cells in a plaque assay, and clones are selected and passaged several times in specific pathogen free (SPF) embryonated chicken eggs.
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