高等学校化学学报

高等学校化学学报 ›› 2004, Vol. 25 ›› Issue (3): 421.

• 研究论文 • 上一篇    下一篇

分子信标荧光探针用于抑癌基因ING1表达产物的定量测定

李军, 王柯敏, 谭蔚泓, 刘斌, 郭秋平, 唐志文, 刘凌风   

  1. 湖南大学生物技术研究院, 化学化工学院, 化学生物传感与计量学国家重点实验室, 长沙 410082
  • 收稿日期:2002-10-28 出版日期:2004-03-24 发布日期:2004-03-24
  • 通讯作者: 王柯敏(1957年出生),男,博士,教授,博士生导师,从事纳米及单分子水平上的生物分析化学及纳米生物技术研究.E-mail;kmwang@mail.hunu.edu.cn E-mail:kmwang@mail.hunu.edu.cn
  • 基金资助:

    国家自然科学基金重点课题(批准号;20135010);国家重点基础研究规划项目(批准号;2002CB513110);海外青年学者合作研究基金(批准号;20028506);国家自然科学青年基金(批准号;200305006);湖南省科技计划重点项目(批准号;03

Quantitative Detection of the Expression Product of Tumor Suppressor Gene ING1 by Molecular Beacon Fluorescence Probe

LI Jun, WANG Ke-Min, TAN Wei-Hong, LIU Bin, GUO Qiu-Ping-TANG Zhi-Wen, LIU Ling-Feng   

  1. Institute of Biological Technology, College of Chemistry & Chemical Engineering, State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082, China
  • Received:2002-10-28 Online:2004-03-24 Published:2004-03-24

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被引次数

28

摘要： 根据抑癌基因ING1基因的序列设计并合成了检测ING1转录产物的分子信标核酸探针,发展了一种快速测量从正常细胞系和鼻咽癌肿瘤细胞系提取的ING1转录产物的方法,所得结果与用逆转录结合PCR法(RT-PCR)得到的结果相吻合.并且,将能表达ING1基因的质粒转入肿瘤细胞进行培养后,再将分子信标转入肿瘤细胞,发现转导了质粒的肿瘤细胞比未转导质粒的肿瘤细胞内的荧光明显增强,从而进一步证实了所设计的分子信标核酸探针与ING1转录产物的结合.

关键词: 分子信标荧光探针, 抑癌基因, 定量测定

Abstract: Three molecular beacons(MBs) were synthesized to find a suitable probe to hybridize the expression of ING1 gene, a recently identified tumor suppressor gene. The loop sequences of MB1 and MB2 are the anti sense and sense sequence of ING1, respectively. The sequence of MB3 is a piece of ssRNA sequence in tobacco mosaic virus, which has no analogy to human gene. MB1 is the most suitable probe because MB1 has the highest fluorescence enhancement after hybridizing with RNA extracted from normal cell. A simple method has been built for the quantitative analysis of ING1 mRNA from nasopharyngral carcinoma(NPC) cell lines and normal cells. The results show that the concentration of ING1 mRNA in normal cells is higher than that in tumor cells. This result is consistent to that of RT PCR detection. ING1 plasmid was delivered into living human NPC cells with liposome and the expression of ING1 by plasmid was monitored by the selected MB.

Key words: Molecular beacon fluorescence probe, Tomur suppresson gene, Quantitative detection

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