赵丽媛,于志刚,甄毓,米铁柱

• 1:中国海洋大学海洋生命学院
• 2:中国海洋大学海洋化学理论与工程技术教育部重点实验室
• 3:中国海洋大学环境与生态教育部重点实验室

摘要(Abstract)：

建立了检测东海原甲藻线粒体细胞色素b(Cytb)基因mRNA含量的实时荧光定量PCR方法。以甲藻Cytb基因的简并引物扩增得到984 bp长的东海原甲藻Cytb基因片段,经克隆、测序,制备并纯化质粒。以光度法测定质粒浓度并作为标准品。对上述基因片段,利用PRIMER EXPRESS 3.0设计引物,以梯度稀释的质粒标准品进行定量PCR反应,以SYBR Green I为荧光染料,制作标准曲线,得到基因拷贝数X与Ct值的关系为:Ct=-3.50 lgX+39.35(相关系数r为0.999)。通过对不同生长时期的东海原甲藻样品的Cytb基因mRNA含量检测,发现该基因的表达量较稳定,平均值为(45.4±4.7)拷贝/细胞,受生长状态影响不大,可作为实时荧光定量PCR的内参基因。

关键词(KeyWords)： 线粒体细胞色素b基因;东海原甲藻;实时荧光定量PCR;内参基因

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(40406028);; 国家高技术发展计划项目(2006AA09Z178)资助

作者(Author): 赵丽媛,于志刚,甄毓,米铁柱

DOI: 10.16441/j.cnki.hdxb.2009.03.015

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