摘要
[目的]研究猪NCOA1基因的PCR-RFLP多态性,为猪的分子育种和标记辅助选择提供理论依据。[方法]以81头山东寿光六和原种猪场的长白猪为研究对象,采取常规酚-氯仿抽提法从猪耳组织提取基因组DNA,以基因组DNA为模板进行PCR扩增获得440 bp的NCOA1基因目的片段,利用限制性内切酶RsaⅠ对PCR产物进行酶切和琼脂凝胶电泳检测,并计算各等位基因频率和基因型频率。[结果]琼脂糖电泳检测酶切产物出现3种基因型:AA型(1条带:440 bp)、AB型(3条带:440、282、158 bp)、BB型(2条带:282、158 bp);基因型频率分别为0.54、0.43和0.03;A和B 2个等位基因的基因频率分别为0.76和0.24。[结论]该研究猪群中,A为优势等位基因,AA基因型频率较高。
[Objective] The research aimed to study the polymorphism of NCOA1 gene in pigs by PCR-RFLP.[Method] Using 81 Landrace pigs from Liuhe pig breeding farm as research objects,the genome DNA was extracted from ear tissue of pigs by conventional Phenol-chloroform extraction method.The genomic DNA was used as template to make PCR amplification and a 440 bp target fragment was obtained.PCR amplification products were digested by restriction nuclease Rsa I.The agarose gel electrophoresis detection was made.And the gene frequency and the genotype frequency were calculated.[Result] The agarose gel electrophoresis detection showed that the enzyme digestion products had three genotypes of AA （440 bp）,AB （440 bp,282 bp,158 bp） and BB （282 bp,158 bp）：The genotype frequencies of AA,AB and BB were 0.54,0.43 and 0.03,respectively：The frequencies of allele A and B were 0.76 and 0.24,respectively.[Conclusion] In the studied pig herd,A is the dominant allele and the genotype frequency of AA is higher.
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~~
