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02 Feb

Influenza A viruses (IAV) pose a constant threat to the human population and therefore a better understanding of their fundamental biology and identification of novel therapeutics is of upmost importance.

Various reporter-encoding IAV were generated to achieve these goals, however, one recurring difficulty was the genetic instability especially of larger reporter genes.

While NS1 is translated from an intron-containing m RNA transcript, the NEP m RNA is generated by exploitation of the cellular splicing machinery (Fig. There, an NS1-GFP fusion protein and NEP are encoded by two non-overlapping genes, which are separated by the porcine Teschovirus-1 2A peptide (PTV-1 2A) coding sequence.

However, we also detected a 40 k Da band corresponding to an NS1-2A-NEP polyprotein with both an NS1- and an NEP-specific antibody in the cell lysate of SC35M-infected cells, indicating that the 2A-mediated “stop-carry on” recoding is not completely efficient.

Based on recent observation that N-terminally GFP-tagged NEP (GFP-NEP) retains its nuclear export and polymerase co-factor function in vitro, we reasoned that reporter genes might be introduced into the viral genome as NEP-fusion constructs.

The development of effective countermeasures, such as vaccines or therapeutics has been complicated by the ability of the viruses to rapidly mutate antigenic determinants or antiviral target structures.

Improved vaccine approaches, identification of new antivirals and in general a better understanding of the fundamental biology of IAV infection is required to efficiently antagonize these human pathogens in future.