Interference 103,781
A. In order to make the shortest possible gene that
we knew made the toxin part of the gene, we knew it would
take over 1800 bases to make it.
And the DNA’s you make on the oligosynthesis machine
are single-stranded, so you’d have to multiply that by two.
Then you’d have to build some overlaps in so they could be
assembled.
. . . . .
Q. And did you make some kind of an estimate of how
long it would take to do that, then, if you were to make an
1800 base pair long DNA?
A. Well, I think it would take me over 500 days if it
was just me working.
Q. And did you discuss the length of that project
with Dr. Adang?
A. Well, Mike and I did talk about that and we
thought if we could get some kind of incremental
improvement, by improving some shorter region of the gene,
such as polyadenylation regions, that that would be an
advantage, because we were really kind of a small company,
and we wanted to have some incremental improvements. We
might be able to go back and make another improvement and
then another improvement and add those on together, to
improve the overall performance of the gene.
Q. What approach did you decide to use for modifying
the gene?
A. We decided to use the approach for looking for a
specific region in the gene to modify. But we agreed that
the way to modify it was to change the codon usage and
balance the ratio of AT to GC, to being more like that of a
plant, and to retain the amino acid sequence of the Bt gene
and change it to the codons preferred by the plant. The
frequencies most commonly used by the plant.
. . . . .
Q. And can you describe for me what you did?
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