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Dunn School: Notebook 1, Oct 1980-Dec 1982

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MS. 12364/2

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Notebook 1, opens with a three page index (table of contents).

This notebook starts with experiments number 1 & 2 on the influenza virus (strain A/NT60/68) in which Bernie Moss and Brownlee were characterizing a DI band (referred to as band 9) by Maxam Gilbert sequencing. Brownlee then moved on to factor IX experiments attempting to sequence the DNA products synthesized by reverse transcriptase from a predicted factor IX oligonucleotide primer (experiment numbers 3-8) and bovine liver mRNA (prepared by Andy Choo, a post-doctoral researcher from Melbourne, Australia). These experiments failed to show any possible factor IX synthetic DNA.

Further experiments relate to the cloning and sequencing of the nucleoprotein band 5 of influenza A/NT/60/68 RNA. Other experiments relate to bands 1-3 of influenza virus A/NT60/68. In particular Brownlee was trying to construct libraries of sonicated or enzyme-restricted ds DNA in m13 phage vectors for Sanger dideoxy sequencing. Construction of new plasmid vector (modified from pAT153) for blunt-ended cloning. New plasmid named pAT153/Pvu II/8 (experiments 55-56). Data sheet prepared, 2 Mar 1982 (see experiment 57, page 119).

In experiments 78-79 and following, e.g. experiment 105, Brownlee attempted cloning of a library of cDNA of polyA+ bovine liver cDNA (in order to isolate factor IX clones) using various enzymatic methods to generate dsDNA and various plasmid cloning vectors. Some clones isolated but inefficient. (See the loose leaf inserted on p. 247- in Andy Choo’s hand – showing batches of polyA+ bovine liver RNA he had prepared - including a purified sucrose gradient fraction. Near the end of the notebook (pages unnumbered) see incomplete notes on a meeting with Dr MP Esnouv on 27 Nov 1980 at the Radcliffe Infirmary. It was at this meeting that Brownlee became aware of Esnouv's work sequencing bovine factor IX. This suggested the approach of first isolating a clone of bovine factor IX since bovine liver was easier to come by than a human liver. Esnouv gave Brownlee some antibodies and bovine factor IX and suggested Brownlee contact Dr Luther at ARC Compton to obtain some calf liver.

Experiment 91 (page 201) concerns a design of a cloning strategy for expression of the influenza haemagglutinin containing clone in mouse fibroblasts.

Dates

  • Oct 1980-Dec 1982

Extent

1 volume

Shelfmark

MS. 12364/2

Repository Details

Part of the Bodleian Libraries Repository

Contact:
Weston Library
Broad Street
Oxford OX1 3BG United Kingdom