Citation: ZHOU Lu, CHENG Hui, WANG Jin-E, PEI Ren-Jun. G-Quadruplex DNAzyme Biosensor for Quantitative Detection of T4 Polynucleotide Kinase Activity by Using Split-to-Intact G-Quadruplex DNAzyme Conversion[J]. Chinese Journal of Analytical Chemistry, ;2016, 44(1): 13-18. doi: 10.11895/j.issn.0253-3820.150480 shu

G-Quadruplex DNAzyme Biosensor for Quantitative Detection of T4 Polynucleotide Kinase Activity by Using Split-to-Intact G-Quadruplex DNAzyme Conversion

1.
Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou 215123, China

Corresponding author: PEI Ren-Jun,
Received Date: 11 June 2015
Available Online: 24 August 2015
Fund Project: 本文系国家自然科学基金(Nos.31301495,21275156,21305154)资助 (Nos.31301495,21275156,21305154)

A biosensor for detection of T4 polynucleotide kinase (PNK) activity was fabricated by using a split-to-intact DNAzyme conversion strategy. G-quadrplex-forming G-rich sequence PS5.M was chosen as a model DNAzyme and splitted into two short parts of S₁OH and S₂OH, which both had hydroxyl groups at 3' and 5'terminals, and could hybridize to another sequence, helper DNA (SH), to form a duplex structure. In the presence of T4 PNK, 5'-end of S₂OH was phophorylated to obtain S₂P. Then S₁OH and S₂P could be linked by T4 DNA ligase to intact PS5.M. In the presence of exonuclease Ⅲ (Exo Ⅲ), SH chain in the duplex of PS5.M-SH could be cleaved into nucleotides, releasing free PS5.M. The released PS5.M could form catalytically active G-quadruplex DNAzyme upon binding hemin in the presence of K⁺, which could catalyze the oxidation of 2,2'-azino bis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) by H₂O₂. Quantitative detection of T4 PNK activity could be achieved by measuring the maximal absorbance of product at 418 nm. The absorbance of the sensing system at 418 nm exhibited a linear relationship with T4 PNK activity in the range of 0.02-0.3 U/mL with a detection limit of 0.014 U/mL (S/N=3). The biosensor was also applied to the determination of PNK activity in Hela cell and HEK 293 cell, and good recoveries of 95.6%-105.7% were obtained at three spiked levels.
- T4 polynucleotide kinase,
- DNAzyme,
- Split-to-intact G-quadruplex,
- Biosensor
1. [1]
  1 Richardson C C. Proc. Natl. Acad. Sci. USA, 1965,54:158-165
2. [2]
  2 Wang L K, Shuman S. Nucleic Acids Res., 2002,30:1073-1080
3. [3]
  3 Cameron V, Uhlenbeck O C. Biochemistry, 1977,16:5120-5126
4. [4]
  4 Wang L K, Shuman S. J. Biol. Chem., 2001,276, 26868-26874
5. [5]
  5 Tang Z, Wang K, Tan W, Ma C, Li J, Liu L, Guo Q, Meng X. Nucleic Acids Res., 2005,33:e97-e97
6. [6]
  6 Song C, Zhao M. Anal. Chem., 2009,81:1383-1388
7. [7]
  7 Lin L, Liu Y, Zhao X, Li J. Anal. Chem., 2011,83:8396-8402
8. [8]
  8 Huang Y, Chen J, Shi M, Zhao S, Chen Z F, Liang H. J. Mater. Chem. B, 2013,1:2018-2021
9. [9]
  9 Hou T, Wang X, Liu X, Pan C, Li F. Sens. Actuators B, 2014,202:588-593
10. [10]
  10 Robertson D L, Joyce G F. Nature, 1990,344:467-468
11. [11]
  11 Breaker R R, Joyce G F. Chem. Biol., 1994,1:223-229
12. [12]
  12 Cuenoud B, Szostak J W. Nature, 1995,375:611-614
13. [13]
  13 SONG Lu-Na, ZHANG Yong-Hua, BO Hong-Yan, GAO Qiang. Chinese J. Anal. Chem., 2015,43(9):1402-1407 宋璐娜, 张永花, 薄红艳, 高强.分析化学,2015,43(9):1402-1407
14. [14]
  14 DUAN Na-Na, WANG Na, YANG Wei, KONG De-Ming. Chinese J. Anal. Chem., 2014,42(10):1414-1420 段娜娜, 王娜, 杨薇, 孔德明.分析化学,2014,42(10):1414-1420
15. [15]
  15 Huppert J L. FEBS J., 2010,277(17):3452-3458
16. [16]
  16 Lipps H J, Rhodes D. Trends Cell Biol., 2009,19(8):414-422
17. [17]
  17 Jiang H, Kong D, Shen H X. Biosens. Bioelectron, 2014,55:133-138
18. [18]
  18 Jiang C, Yan C, Jiang J. Anal. Chim. Acta, 2013,766:88-93
19. [19]
  19 Li T, Wang E, Dong S. Anal. Chem., 2010,82(4):1515-1520
20. [20]
  20 Aleman-Garcia M A, Orbach R, Willner I. Chem-Eur J., 2014,20(19):5619-5624
21. [21]
  21 Xiao Y, Pavlov V, Niazov T, Dishon A, Kotler M, Willner I. J. Am. Chem. Soc., 2004,126(24):7430-7431
22. [22]
  22 Travascio P, Li Y, Sen D. Chem. Biol., 1998,5:505-517
23. [23]
  23 Nagatoishi S, Tanaka Y, Tsumoto K. Biochem. Biophys. Res. Commun, 2007,352:812-817
24. [24]
  24 He H Z, Leung K H, Wang W, Chan D S, Leung C H, Ma D L. Chem. Commun, 2014,50:5313-5315
1. [1]
  Yang Meiqing , Lu Wang , Haozi Lu , Yaocheng Yang , Song Liu . Recent Advances of Functional Nanomaterials for Screen-Printed Photoelectrochemical Biosensors. Acta Physico-Chimica Sinica, 2025, 41(2): 100018-0. doi: 10.3866/PKU.WHXB202310046
2. [2]
  Qiaoqiao BAI , Anqi ZHOU , Xiaowei LI , Tang LIU , Song LIU . Construction of pressure-temperature dual-functional flexible sensors and applications in biomedicine. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2259-2274. doi: 10.11862/CJIC.20240128
3. [3]
  Meijin Li , Xirong Fu , Xue Zheng , Yuhan Liu , Bao Li . The Marvel of NAD⁺: Nicotinamide Adenine Dinucleotide. University Chemistry, 2024, 39(9): 35-39. doi: 10.12461/PKU.DXHX202401027
4. [4]
  Ke Zhao , Zhen Liu , Luyao Liu , Changyuan Yu , Jingshun Pan , Xuguang Huang . Functionalized Reflective Structure Fiber-Optic Interferometric Sensor for Trace Detection of Lead Ions. Acta Physico-Chimica Sinica, 2024, 40(4): 2304029-0. doi: 10.3866/PKU.WHXB202304029
5. [5]
  Lin′an CAO , Dengyue MA , Gang XU . Research advances in electrically conductive metal-organic frameworks-based electrochemical sensors. Chinese Journal of Inorganic Chemistry, 2025, 41(10): 1953-1972. doi: 10.11862/CJIC.20250160
6. [6]
  Xingchao Zhao , Xiaoming Li , Ming Liu , Zijin Zhao , Kaixuan Yang , Pengtian Liu , Haolan Zhang , Jintai Li , Xiaoling Ma , Qi Yao , Yanming Sun , Fujun Zhang . Photomultiplication-Type All-Polymer Photodetectors and Their Applications in Photoplethysmography Sensor. Acta Physico-Chimica Sinica, 2025, 41(1): 100007-0. doi: 10.3866/PKU.WHXB202311021
7. [7]
  Jiarong Feng , Yejie Duan , Chu Chu , Dezhen Xie , Qiu'e Cao , Peng Liu . Preparation and Application of a Streptomycin Molecularly Imprinted Electrochemical Sensor: A Suggested Comprehensive Analytical Chemical Experiment. University Chemistry, 2024, 39(8): 295-305. doi: 10.3866/PKU.DXHX202401016
8. [8]
  Min LIU , Huapeng RUAN , Zhongtao FENG , Xue DONG , Haiyan CUI , Xinping WANG . Neutral boron-containing radical dimers. Chinese Journal of Inorganic Chemistry, 2025, 41(1): 123-130. doi: 10.11862/CJIC.20240362
9. [9]
  Xingyuan Lu , Yutao Yao , Junjing Gu , Peifeng Su . Energy Decomposition Analysis and Its Application in the Many-Body Effect of Water Clusters. University Chemistry, 2025, 40(3): 100-107. doi: 10.12461/PKU.DXHX202405074
10. [10]
  Gonglan Ye , Xia Yin , Feng Xu , Peng Yang , Yingpeng Wu , Huilong Fei . Innovations in “Four-in-One” Inorganic Chemistry Education. University Chemistry, 2024, 39(8): 136-141. doi: 10.3866/PKU.DXHX202401071
11. [11]
  Zhexue Lu , Ping Wu , Huihui Li , Libai Wen . 四“味”一体的无机及分析化学课程思政. University Chemistry, 2025, 40(6): 333-340. doi: 10.12461/PKU.DXHX202405196
12. [12]
  Liang TANG , Jingfei NI , Kang XIAO , Xiangmei LIU . Synthesis and X-ray imaging application of lanthanide-organic complex-based scintillators. Chinese Journal of Inorganic Chemistry, 2024, 40(10): 1892-1902. doi: 10.11862/CJIC.20240139
13. [13]
  Mengyang LI , Hao XU , Zhonghao NIU , Chunhua GONG , Weihui ZHONG , Jingli XIE . Highly effective catalytic synthesis of β-amino alcohols by using viologen-polyoxometalate hybrid materials. Chinese Journal of Inorganic Chemistry, 2025, 41(7): 1294-1300. doi: 10.11862/CJIC.20250080
14. [14]
  Conghao Shi , Ranran Wang , Juli Jiang , Leyong Wang . The Illustration on Stereoisomers of Macrocycles Containing Multiple Chiral Centers via Tröger Base-based Macrocycles. University Chemistry, 2024, 39(7): 394-397. doi: 10.3866/PKU.DXHX202311034
15. [15]
  Xin MA , Ya SUN , Na SUN , Qian KANG , Jiajia ZHANG , Ruitao ZHU , Xiaoli GAO . A Tb₂ complex based on polydentate Schiff base: Crystal structure, fluorescence properties, and biological activity. Chinese Journal of Inorganic Chemistry, 2024, 40(7): 1347-1356. doi: 10.11862/CJIC.20230357
16. [16]
  Chen Hu , Yu Wu , Lianqing Chen , Tao Huang , Chunya Li , Lin Li , Bingguang Zhang , Peng Mei . 基于“三位一体四融合”策略的化学实验室安全课程创新与探索. University Chemistry, 2025, 40(8): 25-32. doi: 10.12461/PKU.DXHX202410088
17. [17]
  Rui LIU , Xinjun ZHOU , Tao WANG . Photocatalytic degradation performance of tetracycline by MOF-74-Mn/g-C₃N₄ Z-type heterojunction. Chinese Journal of Inorganic Chemistry, 2025, 41(9): 1796-1804. doi: 10.11862/CJIC.20250033
18. [18]
  Zhongyan Cao , Shengnan Jin , Yuxia Wang , Yiyi Chen , Xianqiang Kong , Yuanqing Xu . Advances in Highly Selective Reactions Involving Phenol Derivatives as Aryl Radical Precursors. University Chemistry, 2025, 40(4): 245-252. doi: 10.12461/PKU.DXHX202405186
19. [19]
  Yonghui ZHOU , Rujun HUANG , Dongchao YAO , Aiwei ZHANG , Yuhang SUN , Zhujun CHEN , Baisong ZHU , Youxuan ZHENG . Synthesis and photoelectric properties of fluorescence materials with electron donor-acceptor structures based on quinoxaline and pyridinopyrazine, carbazole, and diphenylamine derivatives. Chinese Journal of Inorganic Chemistry, 2024, 40(4): 701-712. doi: 10.11862/CJIC.20230373
20. [20]
  Yihui Song , Shangshang Qin , Kai Wu , Chengyun Jin , Bin Yu . 生物化学在高水平创新型药学人才培养中的交叉融合应用——以去甲基化酶LSD1抑制剂的活性评价为例. University Chemistry, 2025, 40(6): 341-352. doi: 10.12461/PKU.DXHX202406018

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(605)
HTML views(65)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142