Point mutations

Point mutations were already available from previous work, AQP1 and AQP10 cloned in an oocyte expression vector (PolI) that contains the following sequences (from 5′ to 3′): the T7 bacterial promoter, the 5′ untranslated region of the Xenopus β-globin gene, a multiple cloning site, the 3′ untranslated region of the Xenopus β-globin gene, a poly A tract, and a linearizing site (Garneau et al., 2015). These constructs were used as template to produce 12 mutants that are listed in Table 1 along with the oligonucleotide primers exploited. The localization of the residues substituted is also shown in Fig. 2 (C and D).

Uppercase letters correspond to a base pair that was mutated to generate a residue substitution, to remove a restriction site or to add a restriction site.

Location of residues that were substituted in AQP1 and AQP10. The cartons were drawn as described in Fig. 1 using the same color code. (A and B) Chimeras. (C and D) Point substitutions.