Interference 102,728
As to SX 25, Bates No. 483, we find that it is a page in what is said to be a
Research Summary presented by Drs. Singh and Hitzeman on December 14, 1982.
Page 483 is entitled “Future Work” and reads as follows:
Future Work
1. Determination of processing signals.
A. Removal of sequences from the interferon D
expression plasmid - M13 cloning, site-directed
deletion mutagenesis.
B. Construction of expression plasmids with
restriction site followed by the alpha-factor
“pre-pro” sequence.
C. Use of an aminopeptidase?
2. Portable "-factor signals sequence for use with
another promoter.
3. Construction of a promoter fragment [undated, handwritten notation
which states: “from "-factor gene”]
4. Cellular location of non-secreted heterologous proteins.
5. Determination of mRNA levels.
6. Screening of strains for better secretion.
Here, although presented two weeks after Dr. Singh ordered the 24-mer, we find
no mention of the oligonucleotide, loop deletion mutagenesis, or a plan to delete the
nucleotide sequence encoding the glu-ala residues of the " factor spacer sequence
from the interferon D expression plasmid. The “Future Work” does not indicate which
sequences are to be removed from the interferon D expression plasmid, and the only
technique mentioned with respect to the removal of the undisclosed sequences is site-
directed mutagenesis.41 Thus, we find the Research Summary insufficient to
41 As discussed on pp. 38-39 and footnote 40, above, Drs. Singh and Hitzeman
have both testified that site-directed mutagenesis is a different technique. SR 168,
69
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