Interference 103,579
by restriction and cDNA probes, where the 5' and 3' end
of the GBSS gene has been determined more accurately
(Fig. 2). Sequence determination according to Sanger
et al, 1977 of the GBSS gene has been made on subclones
from pSw and pSx in M13mp18 and mp19 as well as pUC19
starting around the 5' end (see SEQ ID No. 5).
The promoter region has been determined as a Bg1II[sic
BgIII]-NsiI fragment (see SEQ ID No. 4). Transcription and
translation start has been determined at an overlapping
Bg1II[sic BgIII]-HindIII fragment. The terminator region
has in turn been determined at a SpeI-HindIII fragment.
The specification of Hofvander’s involved application further
teaches that “[t]he GBSS gene fragments according to the
invention (see SEQ ID Nos 1, 2 and 3, and Fig. 2) were determined
in the following manner . . .” (HR 282, l. 26-28):
The restriction of pSw with NsiI and HindIII gives
fragment I (SEQ ID No. 1) which subcloned in pUC19 is
called 19NH35. Further restriction of 19NH35 with HpaI-
SstI gives a fragment containing 342 bp of the gene
according to the invention. This fragment comprises
leader sequence, translation start and the first 125 bp
of the coding region.
The restriction of pSm with HpaI and NsiI gives
fragment II (SEQ ID No. 2) which subcloned in pJRD184
(Heusterpreute et al, 1987) is called pJRDmitt. Further
restriction of pJRDmitt with HpaI-SstI gives a fragment
containing 2549 bp of the GBSS gene according to the
invention. This fragment comprises exons and introns
from the middle of the gene.
The restriction of pSx with SstI and SpeI gives
fragment III (SEQ ID No. 3) which subcloned in
pBluescript (Melton et al, 1984) is called pBlue3'.
Further restriction of pBlue3' with BamHI-SstI gives a
fragment containing 492 bp of the GBSS gene according
to the invention. This fragment comprises the last
intron and exon, translation end and 278 bp of trailer
sequence.
-34-
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