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

CORC > 苏州纳米技术与纳米仿生研究所 > 中国科学院苏州纳米技术与纳米仿生研究所 > 纳米生物医学与安全研究部 > 裴仁军团队

	G-Quadruplex DNAzyme Biosensor for Quantitative Detection of T4 Polynucleotide Kinase Activity by Using Split-to-intact G-Quadruplex DNAzyme Conversion
	Zhou, L(周璐); Cheng, H(程慧); Wang, JE(王金娥); Pei, RJ(裴仁军)
刊名	CHINESE JOURNAL OF ANALYTICAL CHEMISTRY
	2016
卷号	44 期号:1
通讯作者	Pei, RJ(裴仁军)
英文摘要	G-quadruplex DNAzymes are peroxidase-like complexes formed by hemin and DNA G-quadruplexes. Many chemical sensors and biosensors have been developed by using this kind of DNAzymes. T4 polynucleotide kinase (PNK), which catalyzes the transfer of the gamma-phosphate from ATP to the 5'-hydroxyl termini of polynucleotides, plays an important role in living systems. To build a simple and convenient method to detect the activity of PNK, a split-to-intact DNAzyme was introduced. For this PNK sensor, G-quadruplex-forming G-rich sequence PS5.M was split into two shorter parts S1OH and S2OH, which hybridized to another sequence, helper DNA. In the presence of T4 PNK, 5'-end of sequence S2OH was phosphorylated to S2P. Then S1OH and S2P were linked by T4 DNA ligase to the intact G-quadruplex, PS5.M. The released PS5.M by exonuclease III formed catalytically active G-quadruplex DNAzyme upon hemin binding 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(-1) with a detection limit of 0.014 U mL(-1) (S/N = 3). The method was further applied to the determination of PNK activity in Hela and HEK 293 cell lines, and good recoveries of 95.6%-105.7% were obtained at three spiked levels.
关键词[WOS]	EXONUCLEASE REACTION ; MUTATIONAL ANALYSIS ; AMPLIFIED DETECTION ; DNA-LIGASE ; ENZYME ; PHOSPHORYLATION ; AMPLIFICATION ; INHIBITION ; BEACONS ; SENSOR
收录类别	SCI ; EI
语种	英语
WOS记录号	WOS:000368611800002
内容类型	期刊论文
源URL	[http://ir.sinano.ac.cn/handle/332007/4531]
专题	苏州纳米技术与纳米仿生研究所_纳米生物医学与安全研究部_裴仁军团队
推荐引用方式 GB/T 7714	Zhou, L,Cheng, H,Wang, JE,et al. 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).
APA	Zhou, L,Cheng, H,Wang, JE,&Pei, RJ.(2016).G-Quadruplex DNAzyme Biosensor for Quantitative Detection of T4 Polynucleotide Kinase Activity by Using Split-to-intact G-Quadruplex DNAzyme Conversion.CHINESE JOURNAL OF ANALYTICAL CHEMISTRY,44(1).
MLA	Zhou, L,et al."G-Quadruplex DNAzyme Biosensor for Quantitative Detection of T4 Polynucleotide Kinase Activity by Using Split-to-intact G-Quadruplex DNAzyme Conversion".CHINESE JOURNAL OF ANALYTICAL CHEMISTRY 44.1(2016).