ABSTRACT
Congenital splay leg is a hereditary disease observed in newborn piglets. The etiology and pathogenetic mechanism of the disorder are still unknown. The gene for cyclin-dependent protein kinase inhibitor 3 (CDKN3) was identified as a potential candidate gene in a differential display experiment. We analyzed the gene on sequence variations and compared its expression in M. biceps femoris between healthy and affected piglets. Comparative sequencing of the coding region of three healthy and four splay leg piglets revealed twelve single nucleotide polymorphisms (SNP) resulting in six amino acid exchanges in the deduced sequences. However, all polymorphisms were observed in healthy as well as in splay leg piglets thus excluding structural differences of the gene as a cause of the disease. Besides full length transcripts, we found a variety of aberrantly transcribed cDNA in clones derived from M. biceps femoris of healthy as well as of splay leg piglets. All alternative transcripts coexist with normal cDNA. Expression analysis revealed a trend towards higher values in M. biceps femoris of splay leg piglets supporting the results obtained from a differential display.
Subject(s)
Cyclin-Dependent Kinases/antagonists & inhibitors , Lower Extremity Deformities, Congenital/veterinary , Swine Diseases/genetics , Amino Acid Sequence , Animals , DNA Primers/genetics , Gene Expression , Lower Extremity Deformities, Congenital/genetics , Molecular Sequence Data , Polymorphism, Single Nucleotide/genetics , SwineABSTRACT
Large individual differences were observed in the abundance of transcripts from the hsp70.2 gene in primary fibroblast cultures sampled from 15 different pigs. While previously described functional promoter variants of this gene can partly account for the high variability of heat-induced increased abundance of transcripts, they are unrelated to the observed highly variable absolute amounts of hsp70.2 transcripts. Comparative sequence analysis revealed an alteration of the 3'-untranslated region (3'-UTR) sequences in these samples. The variant 3'-UTR allele proved to increase the half life of the hsp70.2 mRNA in reporter gene assays. It is suggested that the cellular stress response is significantly affected by the action and interaction of both promoter and 3'-UTR variants of the hsp70.2 gene.
