Appeal No. 2003-0936
Application No. 09/532,806
by assay of a reporter gene which is functionally attached
to each construct. As such, once a starting promoter or
intron sequence is provided, any of a number of different
functional deletion mutants of the starting sequence
could be readily prepared.
As indicated above, deletion mutants of the ZMGRP
promoter also could be randomly prepared and then assayed.
With this strategy, a series of constructs are prepared,
each containing a different portion of the clone (a
subclone), and these constructs are then screened for
activity. A suitable means for screening for activity
is to attach a deleted promoter construct to a selectable
or screenable marker, and to isolate only those cells
expressing the marker protein. In this way, a number
of different, deleted promoter constructs are identified
which still retain the desired, or even enhanced, activity.
The smallest segment which is required for activity is
thereby identified through comparison of the selected
constructs. This segment may then be used for the
construction of vectors for the expression of exogenous
protein.
The specification also generally describes various regulatory
elements (Spec., pp. 18-21), terminators (Spec., p. 21), transit
or signal peptides (Spec., pp. 21-23), marker genes (Spec.,
pp. 23-27), and exogenous genes for herbicide resistance, insect
resistance, environment or stress resistance, disease resistance,
mycotoxin reduction, grain quality, etc. (Spec., pp. 27-61), which
are suitable for use in modifying plant characteristics, and
include citations to prior art and summaries of the state of the
art. The specification thereafter discusses assays which may be
employed to determine levels of expression of new transgenic DNA
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