梁斌,刘戈飞,谢建平,李蕊,宋旭红

• 1:汕头大学医学院分子生物学实验室

摘要(Abstract)：

目的检测子宫颈鳞癌组织中辅酶Ⅱ依赖性视黄醇脱氢/还原酶[NADP(H)-dependent retinol dehydrogenase/reductase,NRDR]及其选择性剪接亚型NRDRB1 mRNA的表达,建立SYBR实时定量聚合酶链反应(RQ-PCR)检测NRDR及其选择性剪接亚型mRNA的方法,为准确、快速分析NRDR、NRDRB1基因表达奠定基础。方法根据子宫颈鳞癌组织中NRDR、NRDRB1基因序列,设计并合成引物,将逆转录-聚合酶链式反应(RT-PCR)扩增的基因片段克隆至pGEM-T easy载体,重组质粒经蓝白筛选、酶切、测序鉴定后,作为阳性克隆标准品,用于标准曲线的制备,由此可定量检测出每一样品模板中NRDR、NRDRB1起始拷贝数。结果阳性克隆标准品制作的标准曲线可显示循环阈值与模板浓度具有良好的线性关系(NRDR:r=0.994,NRDRB1:r=0.997),可用于NRDR及其选择性剪接亚型基因表达的起始拷贝数定量测定。结论 RQ-PCR方法较常规PCR技术更为准确、特异、灵敏,对定量检测及研究NRDR及其选择性剪接亚型表达有积极意义。

关键词(KeyWords)： 宫颈肿瘤;实时定量聚合酶链反应;NRDR;选择性剪接;基因表达

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(31071153);; 高等学校博士学科点专项科研基金(20094402120005)

作者(Author): 梁斌,刘戈飞,谢建平,李蕊,宋旭红

DOI: 10.13267/j.cnki.syzlzz.2011.05.012

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