Dunn School: Notebook 22, Feb 2007-Feb 2008
Notebook covers experiments including:
- Continuation of experiments with karyopherins, now concentrating on co transfecting karyopherins (human, FLAG tagged) with 3P-TAP tagged (e.g. experiment 221) or just PB2-TAP tagged alone.
- Series of experiments asking if karyopherins co-purify specifically with tagged 3P. Answer: possibly yes (experiment 235A), although also present in controls. α3 & α6 seem definitely higher.
- Series of experiments with PB2 –TAP alone suggest (experiment 247) that all karyopherins, excepting α2, bind.
- Various crystallization and co-crystallization trials of 5' and 3' strand of vRNA promoter (with post doctoral researcher Jitka Vedova) (e.g. experiments 217, 248). No results forthcoming.
- Mutagenesis of PB2-TAP(WSN) to K627E or D701N (experiments 250 and 263)
- Test of whether 627 mutant alters karyopherin binding in cotransfecion experiments with PB2-TAP followed by gel analysis of proteins. Possible change. (Experiment 265.)
- Test if D701N mutation in PB2-TAP alters karyopherin binding in cotransfection experiments. α4 probably increased, also possibly α3 & α6 (experiments 271, 275).
- Feb 2007-Feb 2008