Figure 5

Characterization of the erl-1(tm2703) allele. (A) Schematic of erl-1 gene: grey and white boxes indicate exons and 3'UTR respectively. The erl-1 region deleted in tm2703 is marked by black line. Primers used for RT-PCR are shown as black arrows. Blue and cyan arrows indicate primers used to confirm tm2703 deletion. Primers binding outside and inside the deleted region are shown in blue and cyan respectively. (B) Genomic deletion in erl-1(tm2703) was confirmed by genomic PCR using primers that bind outside and inside the deleted region as shown in Figure 5A. (C) erl-1 mRNAs isolated from wild type and erl-1(tm2703) worms were amplified by RT-PCR. PCR was performed with either cDNA (+) or RNA (-) using primers depicted in Figure 4A. The weak band in erl-1(tm2703) RNA only sample (-) likely results from amplification of residual genomic DNA in the RNA preparation, i.e. the size of product corresponds to size of erl-1 genomic region. (D) Western blot analysis shows lack of ERL-1 protein in the strain homozygous for erl-1(tm2703). ERL-1 was detected with affinity purified rabbit α-ERL-1 and blot was re-probed with mouse α-actin as loading control.