过氧化物酶增殖体激活受体γ2转基因小鼠的制作及其影响因素分析

目的分析影响转基因小鼠制作的因素,优化转基因动物制作过程,建立适合本实验室的转基因小鼠制作方法。方法利用显微注射方法制作转基因小鼠,对超数排卵的程序、受精卵的收集、外源基因的显微注射、胚胎移植和术后护理等影响因素都进行了对比研究。结果优化了转基因小鼠的制作过程,减少了影响胚胎移植成功的不利因素,提高了过氧化物酶增殖体激活受体γ2(PPAR-γ2)转基因小鼠的制作效率(2.9%)。结论成功制作了PPAR-γ2转基因小鼠,优化转基因小鼠的制作程序是提高转基因成功率的可行途径。     

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.