Appeal No. 2003-0936
Application No. 09/532,806
provides a ready ability to prepare and test sequence
variants, for example, incorporating one or more of the
foregoing considerations, by introducing one or more
nucleotide sequence changes into the DNA. . . . .
The specification teaches that “the technique of site-specific
mutagenesis is well known in the art, as exemplified by various
publications” (Spec., p. 14). “The preparation of sequence
variants of the selected promoter DNA segments using site-directed
mutagenesis is provided as a means of producing potentially useful
species and is not meant to be limiting as there are other ways in
which sequence variants of DNA sequences may be obtained” (Spec.,
p. 15)(emphasis added). According to the specification (Spec.,
pp. 15-16)(emphasis added):
Examples of such methodologies are provided by U.S.
Patent No. 4,237,224, specifically incorporated herein by
reference in its entirety. A number of template dependent
processes are available to amplify the target sequences of
interest present in a sample, such methods being well
known in the art and specifically disclosed herein below.
One efficient, targeted means for preparing
mutagenized promoters or enhancers relies upon the
identification of putative regulatory elements within
the target sequence. This can be initiated by comparison
with, for example, promoter sequences known to be
expressed in a similar manner. Sequences which are
shared among elements with similar functions or
expression patterns are likely candidates for the
binding of transcription factors and are thus likely
elements which confer expression patterns. Confirmation
of these putative regulatory elements can be achieved
by deletion analysis of each putative regulatory region
followed by function analysis of each deletion construct
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