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作 者:孙泰雷[1] 闫守庆[2] 柏蒙蒙[1] 侯永刚[1] 吴明明[1] 李戈[1] 孙金海[1,2]
机构地区:[1]青岛农业大学动物科技学院,山东青岛266109 [2]吉林大学畜牧兽医学院,吉林长春130062
出 处:《Agricultural Science & Technology》2010年第3期52-53,67,共3页农业科学与技术(英文版)
基 金:Supported by Major Specialized Subject of Transgenic Organism New Variety Breeding(20082X08006-003);National Natural Science Foundation of China(30871778 );Construction Engineering Special Fund for Mountain Tai Scholars of Shandong Province~~
摘 要:[Objective] The aim was to study the polymorphism of CMYA3 gene in the 148 pigs of hybrid offspring of 13/17 Robertsonian translocation pigs [2n = 37,rob (13;17)] intercrossing.[Method] PCR-RFLP method was adopted.[Result] A 507 bp fragment of CMYA3 gene was obtained by PCR amplification,and then amplification product by using restriction nuclease Bsh1236Ⅰ was detected by agarose gel electrophoresis.As a result,both alleles (A and B) of the loci were found in the population.The frequencies of allele A and B were 0.699 and 0.301.The genotype frequencies of AA,AB and BB were 0.615,0.169 and 0.216.The frequencies of allele A and genotype AA were significantly higher than allele B and genotype BB in populations.[Conclusion] The study will provide theoretical basis for molecular breeding and marker-assisted selection of 13/17 Robertsonian translocation pigs.[目的]对13/17罗伯逊易位杂合子猪[2n=36,rob(13;17)]互交后代中的148个体进行CMYA3基因的多态性分析。[方法]采用PCR-RFLP方法分析CMYA3基因在13/17罗伯逊易位猪群中的多态性。用酚-氯仿抽提法从猪耳组织中提取基因组DNA。以基因组DNA为模板进行PCR扩增,反应体系为25μl:dNTPs 2μl,上下游引物各0.5μl,Taq酶1 U,模板DNA1μl,用ddH2O补足至25μl。PCR反应条件为:94℃预变性5 min;94℃变性30 s,53℃退火45 s,72℃延伸30 s,共35个循环;72℃延伸10 min。PCR产物经PCR纯化试剂盒纯化,限制性内切酶Bsh1236 I 37℃酶切过夜后,1%琼脂糖凝胶电泳,检测酶切结果。[结果]通过PCR扩增所有个体均能获得507 bp的目的片段,PCR产物经限制性内切酶Bsh1236I酶切,琼脂糖凝胶电泳结果表明该基因在此群体中具有A和B 2个等位基因,基因频率分别为0.699和0.301,具有AA、AB和BB 3种基因型,基因型频率分别为0.615、0.169和0.216。因此,在该试验猪群中,A为优势等位基因,AA基因型频率最大。[结论]该研究为13/17罗伯逊易位猪的分子育种和标记辅助选择提供了理论依据。
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