多鳞鱚不同组织荧光定量PCR内参基因筛选Screening of Reference Genes for Real-time PCR in Different Tissues from Sillago sihama
张宁,杜文文,王中铎,黄洋,杜涛,董忠典
摘要(Abstract):
【目的】筛选多鳞鱚(Sillago sihama)雌雄不同组织中稳定表达的内参基因。【方法】应用实时荧光定量PCR(qRT-PCR)技术定量分析多鳞鱚snrpd1、rps27、rpl7a、rpl7、cnpb、rps4、rps20、ef1a、ube2、rplp2等10个候选内参基因m RNA在雌、雄个体脑、鳃、性腺、心、肠、肾、肝、肌肉、皮肤、脾、胃等22个组织中的表达水平,通过BestKeeper、NormFinder、GeNorm工具测评10个内参基因表达的稳定性。【结果】10个候选内参基因均可获得特异性的扩增产物和理想的扩增效率,其中Bestkeeper软件计算候选内参基因稳定性由高到低的顺序为:snrpd1=rps27> rpl7a> rpl7>cnpb=rps4> rps20> ef1a> ube2> rplp2;Norm Finder软件分析结果为:rpl7>rpl7a> rps27> rps4> cnpb> ef1a> rps20> ube2> rplp2> snrpd1;Ge Norm软件分析结果为:rpl7/rpl7a> rps4>ef1a> rps27> rps20> cnpb> ube2> rplp2> snprd1。【结论】rpl7、rpl7a、rps27基因的表达稳定性较高,建议选择rpl7和rpl7a共同作为多鳞鱚q RT-PCR研究的内参基因。
关键词(KeyWords): 多鳞鱚;荧光定量PCR;内参基因
基金项目(Foundation): 广东省科技计划项目(2015A020209163、2014A020208117);; 广东省海洋渔业科技推广专项项目(A201708A05);; 广东海洋大学博士启动项目
作者(Author): 张宁,杜文文,王中铎,黄洋,杜涛,董忠典
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