Figure 1.

Isolation ofCalpAandCalpBalleles. (A) Genomic organization of the CalpA locus at cytological position 56D5. The position of the CalpAKG05080 P-element, inserted 948 bp upstream from the CalpA translation initiation site, is indicated. The breakpoints and the sizes of the CalpA808 and the Df(2R)ED3716CG2 deficiencies are shown at the bottom. The boxes represent exons in the gene and in the primary transcripts. (B) RT-PCR detects CalpA mRNA in the w1118 control strain and in the P{SUPor-P}CalpAKG05080 insertion line but not in the CalpA808/Df(2R)ED3716CG2 transheterozygotes. Arrows indicate wild-type genomic and complementary DNA RT-PCR fragments. (C) Western blotting of adult whole fly extracts reveals CalpA protein in the w1118 control strain and in the P{SUPor-P}CalpAKG05080 insertion line but not in the CalpA808/Df(2R)ED3716CG2 transheterozygotes. The lower panel shows loading controls; the arrows mark the positions of CalpA and α-actin bands. (D) Genomic organization of the CalpB locus at cytological position 67D1. The location of the CalpBEY08042 P-element, inserted 175 bp upstream from CalpB ATG, is indicated. The breakpoints and the sizes of the CalpB505 and CalpB361 deficiencies are shown at the bottom together with the size of the CBG26 genomic DNA fragment used for the rescue of the mutant phenotypes. The boxes indicate the exons in the primary transcript. (E) RT-PCR reveals the presence of truncated CalpB mRNA in the CalpB505 and CalpB361 homozygotes. The calculated sizes of the PCR products are designated by the arrows on the right. (F) No CalpB-specific signals can be detected in the extracts from adult CalpB505 homozygous mutants by Western blotting. The arrows denote the positions of the full length 110 kDa CalpB in the upper panel, and the α-actin loading control bands in the lower panel, respectively.

Kókai et al. BMC Developmental Biology 2012 12:20   doi:10.1186/1471-213X-12-20
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