TABLE 2.

Overlap extension PCRs for construction of chimeric fiber proteins

ReactionConstructTemplateaPolymeraseb5′ Primer3′ PrimerProgramAmplified fiber protein fragment (residues)
1Ad37s/Ad5kwt Ad37 virusVentL3737s5k-31Ad37 (1-184)
2Ad37s/Ad5kpDV67Vent5k-5fiber32Ad5 (400-581)
3Ad37s/Ad5kRxns 1 and 2 Taq L37fiber31Ad37 (1-184); Ad5 (400-581)
4Ad5s/Ad37kpDV67Ventfiber55s-32Ad5 (1-405)
5Ad5s/Ad37kwt Ad37 virusVent5s37k-537fr2Ad37 (188-365)
6Ad5s/Ad37kRxns 4 and 5 Taq fiber537fr1Ad5 (1-405); Ad37 (188-365)
7Ad5ΔspDV67Ventfiber5short32Ad5 (1-94)
8Ad5ΔspDV67Ventshort5fiber32Ad5 (317-581)
9Ad5ΔsRxns 7 and 8 Taq fiber5fiber31Ad5 (1-94 and 317-581)
10Ad5s/Ad37spDV67Ventfiber5rep3-32Ad5 (1-75); Ad37 (74-89)
11Ad5s/Ad37spDV67Ventrep3-5rep7-32Ad5 (95-370); Ad37 (74-89 and 166-171)
12Ad5s/Ad37spDV67Ventrep7-5fiber32Ad5 (387-581); Ad37 (166-171)
13Ad5s/Ad37sRxns 11 and 12 Taq rep3-5fiber32Ad5 (95-370 and 387-581); Ad37 (74-89 and 166-171)
14Ad5s/Ad37sRxns 10 and 13 Taq fiber5fiber31Ad5 (1-75, 95-370, and 387-581); Ad37 (74-89 and 166-171)
  • a wt, Wild type; Rxns, reactions.

  • b Vent, vent DNA polymerase (New England Biolabs) Taq, Taq DNA polymerase (Gibco BRL).