Questions the researchers asked...hmmmm???

How can we generate mutant M2 strains?

Plasmid construction

Plasmid driven genetics

Chimeric M2 mutants

Will these mutants possess ionic activity?

Two electrode voltage clamp procedure

Do these M2 mutants replicate?

Plaque assay (cell) - Point mutation M2 mutants

Plaque assay (cell) - Chimeric M2 mutants

Do these M2 mutant viruses grow in tissue?

Plaque assay (tissue) Point mutation

Plaque assay (tissue) Chimeric mutants

Amantadine sensitivity test

Does increased M2 incorporation in virions allow for M2 mutant replication?

Western blot

Do these M2 mutant viruses replicate in mice?

Plaque assay

Quiz