Mutation detection in SLC35A3. (A) The deduced amino acid sequence of bovine SLC35A3 and comparison with its homologs in human (Homo sapiens; BAA77841), dog (Canis familiaris; AAC39260), mouse (Mus musculus; AAH24110), and frog (Xenopus laevis; CAD47803). Dots indicate residues that match the Bos taurus sequence. Dashes indicate gaps that have been introduced to optimize the alignment. The valine at position 180 substituted by phenylalanine in CVM is indicated in bold type and shaded in gray. (B) Electropherograms showing the nucleotide sequences across the G→T mutation (indicated by an asterix) in normal (+/+), carrier (+/-), and CVM-affected (-/-) animals. (C) Northern blot analysis of SLC35A3 mRNA. Poly(A)+ RNA was isolated from kidney tissue of CVM calves and unaffected calves and analyzed by hybridizing Northern blots with a 32P-labeled SLC35A3 probe. Subsequently, the membranes were stripped and reprobed with a 32P-labeled GAPDH (glyceraldehyde-3-phosphate dehydrogenase) fragment.
