[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 ...[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.展开更多
鸡核受体辅激活蛋白1基因(NCOA1)mRNA的表达对性腺的发育、成熟均起关键作用,并且与繁殖性状相关。为了建立荧光定量PCR技术检测鸡NCOA1表达量的方法,根据GenBank中NCOA1基因序列设计合成了引物,以β-action基因为内参基因,对荧光定量PC...鸡核受体辅激活蛋白1基因(NCOA1)mRNA的表达对性腺的发育、成熟均起关键作用,并且与繁殖性状相关。为了建立荧光定量PCR技术检测鸡NCOA1表达量的方法,根据GenBank中NCOA1基因序列设计合成了引物,以β-action基因为内参基因,对荧光定量PCR方法进行了评估。结果表明:构建的标准曲线线性关系良好,建立的NCOA1基因荧光定量PCR检测方法灵敏度高、特异性强,准确可靠。建立的SYBR Green I实时荧光定量PCR方法可以测定鸡NCOA1mRNA表达水平的相对含量。展开更多
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~~
文摘[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.
文摘鸡核受体辅激活蛋白1基因(NCOA1)mRNA的表达对性腺的发育、成熟均起关键作用,并且与繁殖性状相关。为了建立荧光定量PCR技术检测鸡NCOA1表达量的方法,根据GenBank中NCOA1基因序列设计合成了引物,以β-action基因为内参基因,对荧光定量PCR方法进行了评估。结果表明:构建的标准曲线线性关系良好,建立的NCOA1基因荧光定量PCR检测方法灵敏度高、特异性强,准确可靠。建立的SYBR Green I实时荧光定量PCR方法可以测定鸡NCOA1mRNA表达水平的相对含量。