分子信标用于核酸连续复制过程的体外实时监测

高等学校化学学报 ›› 2009, Vol. 30 ›› Issue (8): 1538.

分子信标用于核酸连续复制过程的体外实时监测

孟祥贤, 羊小海, 王柯敏, 郭秋平, 李军, 唐志文

湖南大学化学生物传感与计量学国家重点实验室, 化学化工学院, 生物医学工程中心, 生物纳米与分子工程湖南省重点实验室, 长沙 410082

收稿日期:2009-03-10 出版日期:2009-08-10 发布日期:2009-08-10
通讯作者: 王柯敏, 男, 博士, 教授, 博士生导师, 主要从事纳米及分子水平上的生物分析化学及纳米生物技术研究. E-mail: kmwang@hnu.cn
基金资助:
国家自然科学基金(批准号: 20505007, 90606003)和湖南省自然科学基金(批准号: 08JJ1002) 资助.

Real-time Monitoring Continuous Replication of DNA Leading Strand in vitro Based on Molecular Beacon

MENG Xiang-Xian, YANG Xiao-Hai, WANG Ke-Min^*, GUO Qiu-Ping, LI Jun, TANG Zhi-Wen

State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Biomedical Engineering Center, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University, Changsha 410082, China)

Received:2009-03-10 Online:2009-08-10 Published:2009-08-10
Contact: WANG Ke-Min. E-mail: kmwang@hnu.cn
Supported by:
国家自然科学基金(批准号: 20505007, 90606003)和湖南省自然科学基金(批准号: 08JJ1002) 资助.

摘要/Abstract

摘要：

利用分子信标核酸探针实时监测了核酸连续复制过程. 分子信标不仅作为模板参与复制反应, 而且同步将复制过程的信息转换为荧光信号, 实现复制过程的体外实时监测. 该方法不仅为DNA复制检测提供了一种实时研究手段, 而且为核酸复制动力学及与复制相关疾病的深入研究提供了一种新的思路.

关键词: 分子信标; 核酸复制; 实时监测; 荧光信号

Abstract:

Real-time monitoring of replication of DNA leading strand using molecular beacon in vitro was reported. DNA replication plays a pivotal role in life process. DNA replication is usually assayed by denatured gel electrophoresis and autoradiography, which are complex, time-consuming, and incapable of acquiring the dynamic data in real time. We employed molecular beacon as the probe and template for DNA replication and converted directly replication information into fluorescence signal. The replication information of DNA leading strand was obtained conveniently and accurately using molecular beacon. This method would provide a strategy for real-time monitoring DNA replication and for further research of replication dynamic data and some deseases associated with DNA replication.

Key words: Molecular beacon; DNA replication; Real-time monitoring; Fluorescent signal

TrendMD:

孟祥贤, 羊小海, 王柯敏, 郭秋平, 李军, 唐志文. 分子信标用于核酸连续复制过程的体外实时监测. 高等学校化学学报, 2009, 30(8): 1538.

MENG Xiang-Xian, YANG Xiao-Hai, WANG Ke-Min*, GUO Qiu-Ping, LI Jun, TANG Zhi-Wen. Real-time Monitoring Continuous Replication of DNA Leading Strand in vitro Based on Molecular Beacon. Chem. J. Chinese Universities, 2009, 30(8): 1538.

参考文献

[1]Watson J. D., Crick F. H.. Nature[J], 1953, 171(4361): 964—967
[2]Watson J. D., Crick F. H.. Nature[J], 1953, 171(4356): 737—738
[3]Meselson M., Stahl F. W.. Proc. Natl. Acad. Sci. USA[J], 1958, 44: 671—682
[4]Okazaki R., Okazaki T., Sakabe K.. Proc. Natl. Acad. Sci. USA[J], 1968, 59: 598—605
[5]CHEN Yue-Zeng(陈阅增). General Biology(普通生物学)[M], Beijing: Higher Education Press, 2001: 412—436
[6]Berndt M., Jane B., Carmen F., et al.. J. Cell Biol.[J], 2007, 179: 1385—1398
[7]Tonatiuh R. S., Irina P., Olga L., et al.. Nucleic Acids Res.[J], 2009, 37(1): 38—46
[8]Tsurimoto T., Stiillman B.. J. Biol. Chem.[J], 1991, 266(3): 1950—1960
[9]Minden J. S., Marians K. J.. J. Biol. Chem.[J], 1985, 260(16): 9316—9325
[10]Huang T., Chen C. W.. J. Bacteriol.[J], 2008, 190: 755—758
[11]Toshiki T., Bruce S.. J. Biol. Chem.[J], 1991, 266(33): 1950—1960
[12]Hellman A., Rahat A., Scherer A. W., et al.. Mol. Cell. Biol.[J], 2000, 20: 4420—4428
[13]Nishio M., Ohtsuka J., Tsurudome M., et al.. J. Virol.[J], 2008, 82: 6130—6137
[14]Ono H. A., Le L. P., Davydova J. G., et al.. Cancer Res.[J], 2005, 65: 10154—10160
[15]Tyagi S., Kramer F. R.. Nat Biotech.[J], 1996, 14: 303—306
[16]Fang X., Li J. J., John P., et al.. Anal Chem.[J], 2000, 72: 747A—753A
[17]MENG Xiang-Xian(孟祥贤), WANG Ke-Min(王柯敏), TAN Wei-Hong(谭蔚泓), et al.. Chinese Sci. Bull.(科学通报)[J], 2003, 48: 2132—2136
[18]Vitko J., Rujan I., Androga L., et al.. Biophys. J.[J], 2007, 93(9): 3210 — 3217
[19]Lin Y. W., Ho H. T., Huang C. C., et al.. Nucleic Acids Res.[J], 2008, 36(19): e123
[20]Chen A. K., Behlke M.A., Tsourkas A.. Nucleic Acids Res.[J], 2008, 36(12): e69
[21]LIU Ling-Feng(刘凌风), TANG Zhi-Wen(唐志文), WANG Ke-Min(王柯敏), et al.. Chem. J. Chinese Universities(高等学校化学学报)[J], 2003, 24(10): 1761—1764
[22]Li J. J., Chu Y., Lee B. Y., et al.. Nucleic Acids Res.[J], 2008, 36(6): e36
[23]Horejsh D., Martini F., Poccia F., et al.. Nucleic Acids Res.[J], 2005, 33(2): e13
[24]Li J. J., Geyer R., Tan W. H., et al.. Nucleic Acids Res.[J], 2000, 28(11): e52
[25]Waltz T. L., Marras S., Rochford G., et al.. J. Clin. Microbiol.[J], 2005, 43: 254—258
[26]TANG Zhi-Wen(唐志文), WANG Ke-Min(王柯敏), TAN Wei-Hong(谭蔚泓), et al.. Chinese Sci. Bull.(科学通报)[J], 2003, 48: 1030—1035
[27]Tang Z. W., Wang K. M., Tan W. H., et al.. Nucleic Acids Res.[J], 2003, 31(23): e148
[28]Tang Z. W., Wang K. M., Tan W. H., et al.. Nucleic Acids Res.[J], 2005, 33(11): e97
[29]MENG Xiang-Xian(孟祥贤), TANG Zhi-Wen(唐志文), WANG Ke-Min(王柯敏), et al.. Chinese Sci. Bull.(科学通报)[J], 2006, 51: 2733—2737