Interference No. 102,572
1* when he digested plasmid pBR322(Xap) with EcoRI, filled in, digested with PstI, and
purified by PAGE. He isolated a 1543 bp fragment by treating pGammaCEAInt2 with PstI
followed by BamHI and purification by PAGE. He also isolated a 869 bp fragment from
pGammaCEAInt2 by digestion with AvaI, filling in, cleaving with BamHI and subsequent
PAGE purification. He then ligated these fragments, transformed the ligation reaction into
E. coli and analyzed the resultant colonies by restriction analysis to confirm
13
pGammaCEAtrp207-1*. Holmes, ¶ 19
(CR-31-32).
(13) Accordingly to Cabilly he transformed competent E. coli cells with
pKCEAtrp207-1*delta and retransformed the successful E.coli cells with
pGammaCEAInt2 which confers resistance to ampicillin but not to tetracycline.
Cabilly, ¶ 6 (CR-39-40). He grew the cotransformed cells in minimal media containing
ampicillin and tetracycline and induced the cultures with indoleacrylic acid (IAA) to make
refractile body preparations. Cabilly testified that he gave a sample to Jeanne Perry for
SDS-PAGE analysis. Cabilly indicated that he analyzed several samples by SDS-PAGE;
subsequently these were silver stained or subjected to Western blot using anti-mouse IgG
14
for the identification of light and heavy chain protein. Cabilly, ¶ 7 (CR-
13The Cabilly et al. brief (page 13) argues that this work was done by December 8,
1982. This argument is also not supported by any testimony. id.
14The Cabilly et al. brief (page 14, sec. 4) alleges that the Western Blot used
125
rabbit anti-mouse primary antibodies and I-labeled protein A. This is attorney argument.
16
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