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Molecular genetics of the apolipoprotein B gene in pigs in relation to atherosclerosis
Authors:Maeda Nobuyo   David L. Ebert   Thomas M. Doers   Marsha Newman   Judith Hasler-Rapacz   Alan D. Attie   Jan Rapacz  Oliver Smithies  
Affiliation:Laboratory of Genetics, University of Wisconsin, Madison.
Abstract:Immunologically defined alleles of the pig apolipoprotein B (ApoB) locus (apoB) are correlated with different blood cholesterol levels and predisposition towards premature coronary heart disease. We show here that these alleles are associated with differences in the apoB gene by identifying six restriction fragment length polymorphisms at the pig apoB locus. We have sequenced a 2.4-kb fragment encompassing exons 11 through 14 of one allele, and 7.1 kb from the 3' one-third of exon 26 to about 1 kb past the last exon from another allele. The decoded amino acids of the pig and human ApoB proteins are identical at 70% of these positions. One region close to the C-terminus of the protein is surprisingly different in pigs and humans (57% identity) but the C-terminal region is relatively well conserved (74% identity). Neither of the two putative low-density lipoprotein (LDL) receptor-binding sites is completely conserved in pigs and humans, but identical stretches of amino acids occur near these sites more frequently than in the other sequenced regions. We compare the nucleotide sequences of the region encompassing the putative LDL receptor-binding sites from four pig alleles, including one implicated directly in atherosclerosis. None of the differences appears to account for the hypercholesterolemic phenotype. We conclude that significant differences in the physiology of LDL particles result from changes outside the putative receptor-binding region.
Keywords:Recombinant DNA   nucleotide sequence   RFLP   allelic differences   exon   intron   λ   Charon libraries   LDL receptor binding sites   hypercholesterolemia
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