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Fig. 3. Isolation of the CUP gene. (A) Only one of the isolated
Antirrhinum NAC-domain genes belonged to the same clade as CUC1,
CUC2 and NAM (shaded). Other Arabidopsis NAC-domain
proteins are included for comparison. Accession numbers are: NAP, CAA10955;
CUC1, BAB20598; CUC2, BAA19529; NAM, CAA63101; CUP, AJ568269; GRAB2, CAA09372;
CUC3, AAP82630; AtNAC1, AAF21437; and GRAB1, CAA09371. The tree was
constructed by the neighbour-joining method in MacVector, using only the
conserved NAC-domains. Bootstrap values are indicated. (B) The CUP
gene consists of three exons. Untranslated transcribed regions are shown in
grey and the conserved NAC-domain is stippled. The transposon insertion sites
are shown for each allele. DNA sequence analysis of PCR products spanning the
insertion sites indicated that the transposons were related to Tam-1
(cup-1) (Nacken et al.,
1991) and Tam-4 (cup-2)
(Luo et al., 1991),
respectively. (C) Northern blot hybridised with a truncated CUP
probe, lacking the NAC-domain, shows expression in the wild-type control but
not in the two cup alleles. Stripping and re-hybridisation of the
northern with a constitutive GAPDH probe demonstrates that RNA is
present in each lane.
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