Interference 103,579
length genomic DNA sequences coding for PGBSS in the antisense
direction versus the experimental results for fragments of full
length genomic DNA sequences coding for PGBSS in the antisense
direction reported in Kuipers’ 1995 publication (VDX 4, p. 752,
col. 2; emphasis added):
In transgenic clones, the degree of inhibition of GBSS gene
expression was found to vary for the genomic GBSS antisense
constructs. However, similar frequencies of complete and
incomplete inhibition could be achieved with pGBA10, pKGBA10
and pKGBA31 (comprising 0.6kb of the 3' end of the GBSS
coding region and containing one intron sequence). This
indicates that the size of the antisense RNA does not affect
the efficacy of inhibition. Furthermore, it demonstrates
that the GBSS fragment used in pKGBA31, or at least part of
it, is essential for the inhibition of GBSS gene expression,
as the inhibitory effect of pGBA20, pKGBA20 and pKGBA25 was
much lower.
For pGBA30 and pKGBA30, the weak inhibitory effect
may be caused by a premature transcription termination.
The genomic fragment used for these constructs contains a
3' non-GBSS sequence, which comprises a part of a putative
pseudogene (van der Leij et al. 1993), in addition to the
GBSS fragment that is also present in pKGBA31. . . . A
premature transcription stop does not necessarily result
in the absence of antisense inhibition, as has been
described for pGB50 (Kuipers et al. 1994) and several
other antisense genes . . . but in the case of pGBA30 and
pKGBA30 the resulting antisense RNA might lack sequences
that are complementary to the GBSS mRNA.
We find from the factual evidence in Table 1 of Kuipers 1995
publication (VDX 4, p. 748, Fig.1A,B and Table 1), Visser’s
involved application (VR 139+), and Hofvander’s involved
application (HR 275+, especially HR 312, Fig. 2):
-112-
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