非标记型p53抑癌基因的荧光DNA生物传感器-陕西师范大学学报期刊社网站

陕西师范大学学报（自然科学版）

化学与材料科学

非标记型p53抑癌基因的荧光DNA生物传感器

刘萍1，2，白金燕1，金燕1*

(陕西师范大学化学与材料科学学院，陕西西安 710062；2 商洛学院化学与化学工程系，陕西商洛 726000)

刘萍，女，讲师，主要研究方向为生物传感器和生命分析.E-mail:liupingchenghui@126.com.* 通信作者：金燕，女，副教授，博士.E-mail: jinyan@snnu.edu.cn.

摘要:

基于发夹型核酸探针的高特异性识别能力以及结晶紫与单、双链DNA结合后荧光强度的差异，以发夹型核酸作为探针，构建了一种非标记型检测p53抑癌基因的荧光DNA生物传感器.此传感器不但能够区分完全互补、单碱基突变和随机目标DNA序列，而且还能识别不同突变位点的单碱基突变序列.在0.05～5.00 μmol/L范围内，完全互补目标DNA的浓度与体系的荧光增强效率呈线性关系，检出限为5×10-8mol/L.该方法已应用于检测正常和单碱基突变的p53基因.

关键词：

发夹型核酸探针；结晶紫； p53基因；荧光DNA生物传感器

收稿日期：

2010-04-14

中图分类号：

O657.3

文献标识码：

文章编号：

1672-4291(2011)02-0045-05

基金项目：

国家自然科学基金资助项目（20705017）.

Doi:

Label-free fluorescence DNA biosensor for detection of p53 tumor suppressor gene

Liu Ping1,2,Bai Jin-yan1, Jin Yan1*

(1 College of Chemistry and Materials Science, Shaanxi Normal University, Xi′an 710062, Shaanxi, China; 2 Department of Chemistry and Chemical Engineering, Shangluo University, Shangluo 726000, Shaanxi, China)

Abstract:

A label-free fluorescence biosensor has been developed for detection of p53 tumor suppressor gene based on the specific recognition of hairpin DNA and fluorescence difference derived from interactions of crystal violet (CV) with the ssDNA and dsDNA. The results indicated that this method can easily discriminate complementary target DNA from single-base mutation oligonucleotide and random oligonucleotide. And it also could distinguish the mutation site of single-base mutation oligonucleotide. In the range of 0.05-5 μmol/L，the concentration of complementary sequence was linear with the change of the fluorescence signal of CV, and the detection limit was 50 nmol/L. This method can be used in detecting normal and single-base mutation p53 gene.

KeyWords:

hairpin DNA; crystal violet; p53 gene; fluorescence DNA biosensor