RESEARCH COMMUNICATIONS OF THE 28th ECVIM-CA CONGRESS

A DM‐associated haplotype (c.‐227A, c.‐378G, c.‐420G, c.‐452G; p=0.007) was identified within the putative promoter of ACP1 in a case‐control association study (residual blood samples used for PCR). A dual‐luciferase reporter assay was used to investigate the potential impact of these SNPs on gene expression. DNA sequences carrying two variant promoter haplotypes (variant 1 ‐ DM‐associated haplotype, variant 2 – non‐diabetic haplotype) were amplified by PCR, and cloned separately into the pGL4 vector containing Firefly luciferase [FLuc]. The pRL_CMV vector, containing Renilla luciferase [RLuc] was used as a control vector for co‐transfection of CHO cells, to enable normalisation of FLuc activity. Positive (pGL3) and negative (promoterless pGL4) controls were included. There was no difference in the normalised luminescence comparing the recombinant plasmids (FLuc:RLuc: mean (SE); variant 1: 5.6 (0.42) ; variant 2: 5.13 (0.27) ; p=0.7 [Mann‐Whitney‐U]).