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1、荧光增强法用于解螺旋酶的检测荧光增强法用于解螺旋酶的检测陈陈敬敬华华1， ，张张静静2，李光文，李光文1， ，罗红罗红斌斌1，林清，林清钰钰1， ，陈陈梅梅11福建医科大学 药学院 药物分析系，福建 福州 3500042福建农林大学 生命科学学院 化学生物学系，福建 福州 350002*Email: cjh_设计含有特定碱基对的双螺旋 DNA 片段，DNA 一条链的 5端标记荧光发射基团，另一条链 3末端的 鸟嘌呤（G）碱基作为猝灭基团。解旋酶不存在时，双螺旋结构稳定存在，由于荧光共振能量转移，此时 荧光猝灭。解旋酶存在时，由于酶对 DNA 的特异性解旋作用，双链 DNA 解离，使末端标记的荧
2、光基团和 G 碱基远离，从而使体系的荧光增强。通过解旋酶加入前后体系荧光信号的变化，实现对解旋酶的检测。 结果表明，荧光增强值随着解旋酶浓度的增大而增大，在最佳条件下，该体系可以用于解旋酶的测定。在 1 nM0.5 M 范围内，荧光强度与解螺旋酶的浓度呈良好的线性关系，检测限达 0.3 nM。本文提出了检测 解旋酶的新策略。同时利用解旋酶的解旋特性，可以设计一种新型的 DNA 生物传感策略，课题组目前也 正将其用于乳腺癌相关基因的检测，有望研制一种新型的 DNA 生物传感器用于乳腺癌的早期诊断。 关键词：荧光增强法；丙肝解旋酶；DNA 参考文献 1 Ha T, Rasnik I, Cheng
3、W, Babcock H P, Gauss G H, Lohman T M, Chuc S H. Nature, 2002, 419: 638. 2 Theissen B, Karow A R, Kahler J, Gubaev A, Klostermeier D. Proc. Natl. Acad. Sci. USA, 2008, 105: 548.The Fluorescence Enhancement Method for Detection of HelicaseJinghua Chen1, Jing Zhang2, Guangwen Li1 Hongbin Luo1, Qingyu
4、LIN1, Mei Chen1 1 Department of Pharmaceutical Analysis, Fujian Medical University, Fuzhou 3500042 School of life sciences, Fujian Agriculture and Forestry University, Fuzhou 350002A double helix containing specific base pairs was designed. A complementary strand with appropriate length and rich gua
5、nine bases at 5-end was introduced to form duplex structure with the 3-end fluorophore-labeled strand. In the absence of helicase, fluorescence of fluorophore is quenched by nearby guanine bases. In contrast, the binding of helicase and duplex will destruct the duplex and dissociate the duplex, whic
6、h separates fluorophore from guanine bases and increases the fluorescence signal. Thus, a new continuous fluorescence assay method was developed based on fluorescence resonance energy transfer (FRET) for the monitoring of helicase activity. The result indicated that fluorescence intensity increased
7、with helicase concentration. Therefore, in the best condition, this system can be used in the determination of helicase. It was found that the fluorescence intensity showed a fine linear relationship with the concentration of helicase in the range from 1 nM to 0.5 M. The detection limit (3/S) for he
8、licase was calculated to be 0.3 nM. A new detection strategy for the breast cancer by spiral enzyme was introduced in our study. It is expected to provide a new DNA biosensor for early diagnosis of breast cancer.基金项目：国家自然科学基金 21105012, 福建省高校杰出青年科研人才培育计划 JA11105、JA10295, 福建省自然科学基金 2011J01028, 福建省医学创新基金 2011-CX-22, 福建省教育厅基金资助项目 (JA09116)；福建省大学生创新性实验计划 (34)
