First evidence of host range expansion in virophages and its potential impact on giant viruses and host cells

Said Mougari; Nisrine Chelkha; Dehia Sahmi-Bounsiar; Fabrizio Di Pinto; Philippe Colson; Jonatas Abrahao; Bernard La Scola

The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/780841 doi: bioRxiv preprint A specific PCR system targeting the collagen-like protein (ORF8) was designed and performed on the DNA extracted from supernatant containing the mutant genotype to confirm the occurrence of deletion of the 81 nucleotide-long sequence in this strain (Fig. 3a) . The details of the primers used are listed in Table 1 , and the PCR product was visualized on an agarose 2% gel using SYBR safe buffer (Invitrogen, USA). The Qiagen gel extraction kit was used to recover the band corresponding to each genotype of Guarani detected in Tupanvirus supernatant (wild-type and mutant) from the 2% agarose gel according to the manufacturer's instructions. Sanger sequencing analyses were then performed in an Applied Biosystems ® 3130/3130xl Genetic Analyzer (Thermo Fisher, USA) using a Big Dye Terminator Cycle Sequencing Kit (Thermo Fisher, USA) according to the manufacturer's instructions.