DETECTING LOW DENSITY LIPOPROTEIN RECEPTOR MUTANT GENE OF RABBIT BY PCR

Objective Watanabe Heritable Hyperlipidaemic （WHHL） rabbits with low density lipoprotein receptor （LDLr） gene mutation have provided unprecedented opportunities for the study of human atherusclerusis, in order to confirm LDL receptor gene status in rabbits, we developed a simple PCR technique to detect LDL mutations in rabbits. Methods Rabbits genomic DNA were extracted from ear biopsy, and amplified by PCR to detect 12 bp deletion mutation in WHHL rabbits. PCR products were directly digested with Bgl Ⅰ, and then applied to polyacrylamide gel electrophoresis. Results PCR products from homozygous LDLr ＋/＋ rabbits generated 2 bands of 212 and 94 bp after Bgl Ⅰ digestion, LDLr ＋/- rabbits generated 3 bands （294, 212, and 94 bp）, LDLr -/- animals, however, generated only 1 product （294 bp）. Conclusion This modified PCR method is simple and reliable.

DETECTING LOW DENSITY LIPOPROTEIN RECEPTOR MUTANT GENE OF RABBIT BY PCR

Objective Watanabe Heritable Hyperlipidaemic (WHHL) rabbits with low density lipoprotein receptor (LDLr) gene mutation have provided unprecedented opportunities for the study of human atherosclerosis, in order to confirm LDL receptor gene status in rabbits, we developed a simple PCR technique to detect LDL mutations in rabbits. Methods Rabbits genomic DNA were extracted from ear biopsy, and amplified by PCR to detect 12 bp deletion mutation in WHHL rabbits. PCR products were directly digested with BglⅠ, and then applied to polyacrylamide gel electrophoresis. Results PCR products from homozygous LDLr +/+ rabbits generated 2 bands of 212 and 94 bp after BglⅠ digestion, LDLr +/- rabbits generated 3 bands (294, 212, and 94 bp), LDLr -/- animals, however, generated only 1 product (294 bp). Conclusion This modified PCR method is simple and reliable.