ViralORFeome: an integrated database to generate a versatile collection of viral ORFs

Pellet, J.; Tafforeau, L.; Lucas-Hourani, M.; Navratil, V.; Meyniel, L.; Achaz, G.; Guironnet-Paquet, A.; Aublin-Gex, A.; Caignard, G.; Cassonnet, P.; Chaboud, A.; Chantier, T.; Deloire, A.; Demeret, C.; Le Breton, M.; Neveu, G.; Jacotot, L.; Vaglio, P.; Delmotte, S.; Gautier, C.; Combet, C.; Deleage, G.; Favre, M.; Tangy, F.; Jacob, Y.; Andre, P.; Lotteau, V.; Rabourdin-Combe, C.; Vidalain, P. O.

In this recombination-based cloning pipeline, viral ORFs amplified by PCR or RT-PCR are cloned by in vitro recombination into donor vectors such as pDONR207 or pDONR223 (see Supplementary Data for detailed protocols). After transformation, a construct can be purified either from a mini-pool of bacteria colonies to keep the sequence diversity of the original template (e.g. when working with viral quasi-species) or from a single isolated colony. These two cloning strategies, referred as 1.0 and 2.0, respectively (5), are manageable using ViralORFeome interface (Figure 2 ).