Dunn School: Notebook 16, Feb 2000-Nov 2000

File

MS. 12364/19

Held in our offsite storage facility

Influenza related experiments:

A NP DI (about 420 nt) giving an abnormally truncated NP of 52 amino acids (experiment 616).
Successful sequencing of eleven independent clones derived from full length band 8 cloning, ranging from 9-17 long mRNA snatched sequences (experiment 620). Not published.
Unsuccessful attempts to detect "hybrid transcripts" with a ppp 5' end and poly (A) 3' end.
Unsuccessful experiments using pppA (32P labelled in γ position) to study cDNA transcripts of 49 mer RNA.
Start of transient transfection (lipofectamine) of 293T cells with 4 cDNA plasmids (for PB1, PB2, PA & NP) to obtain crude recombinant flu polymerase expressed in 293T cells (experiment 633) (experiment 649, very encouraging) (also experiment 641, using GTP label) copying of vRNA 49-mer.
Transient transfection of 293T cells with mutant PA (post doctoral researchers Ervin Fodor and Mandy Crow) (experiment 650).
First experiments (looked hopeful) with short synthetic primers of 5' (15 mer) and 3' end (14 mer) of vRNA promoter and transfected nuclear extracts with GTP labelled transcript on 18% gel (experiment 653).
Heat inactivation studies of PA mutants (from Ervin Fodor and Mandy Crow) of reconstituted recombinant polymerase (experiment 656, 660, 661, 667).
Start of initially unsuccessful experiments to test for endonuclease activity of recombinant polymerase (e.g. experiment 668 with Mike Leahy, post-doctoral researcher).

Dates

1 volume

MS. 12364/19

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