山羊IFN-γ和TNF-α基因SYBR Green Ⅰ实时荧光定量RT-PCR检测方法的建立及应用OA北大核心CSCDCSTPCD
Establishment and application of SYBR Green Ⅰ real-time fluorescence quantification RT-PCR for IFN-γ and TNF-α genes of goat
根据GenBank中山羊γ-干扰素(IFN-γ)和肿瘤坏死因子-α(TNF-α)的基因序列,设计特异引物扩增目的基因.将IFN-γ、TNF-α基因克隆至pMD-19-T载体,得到各自阳性克隆质粒,以2种阳性质粒为标准品建立标准曲线并进行熔解曲线分析以及灵敏性、特异性和重复性试验.应用所建立的方法检测刀豆蛋白A(ConA)刺激健康山羊外周血单核细胞(PBMC)后不同时间点IFN-γ、TNF-α基因转录量.结果表明,当质粒标准品稀释度为1×109 c…查看全部>>
Interferon-γ(IFN-γ) and tumor necrosis factor-α(TNF-α) of goat were selected according to GenBank to design specific primers to amplify the target genes.The two kinds of genes were cloned into pMD-19-T vector to obtain the respective positive clone plasmids,which were used as standards to create a standard curve.Melting curve analysis as well as sensitivity,specificity and reproducibility experiments were conducted.With the proposed method,the gene transcrip…查看全部>>
李婷;张益;鲜思美;包细明;冯将;李鹏飞
贵州大学动物科学学院,贵州贵阳550025贵州省动物疫病研究所,贵州贵阳550025贵州大学动物科学学院,贵州贵阳550025贵州大学动物科学学院,贵州贵阳550025贵州大学动物科学学院,贵州贵阳550025贵州大学动物科学学院,贵州贵阳550025
农业科技
山羊细胞因子实时荧光定量RT-PCRSYBR Green Ⅰ
caprinecytokinesreal-time fluorescence quantitative PCRSYBR Green Ⅰ
《中国兽医科学》 2017 (10)
1292-1298,7
贵州省科技计划课题[黔科合LH字(2014)7667]贵州大学2015年大学生创新创业训练计划项目[贵大(省)创字2015(005)]贵州省动物疫病防控与兽医公共卫生保障科技创新人才团队项目[黔科合人才团队(2015)4016号]
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