Comparative Analysis of e6_e7 Splicing and E7 Translation Efficiency Among Alpha Papillomavirus

This poject goal was to assess i) the diversity of splicing coming from the e6 ORF from multiple genotypes of papillomaviruses from the alpha genus and ii) the impact of splicing on the translation efficiency from the downstream e7 ORF. To study splicing and translation, we designed vectors containing for each genotype the e6 ORF followed by the 5'end of the e7 orf, fused to egfp. After expression in the U-2 OS model, spliced and unspliced heterologous bicistronic e6_5'e7-egfp mRNAs were amplified by RT-PCR then submitted to MinION sequencing to confirm prevoiously discovered alternative spliced isoform and to discover new ones.