基于末端脱氧核苷酸转移酶扩增DNA-铜纳米簇荧光传感检测L-组氨酸

发布时间：2018-02-16 11:55

本文关键词： 末端脱氧核苷酸转移酶聚胸腺嘧啶铜纳米簇 L-组氨酸荧光分析　出处：《分析化学》2017年10期 　论文类型：期刊论文

【摘要】：利用末端脱氧核苷酸转移酶(TdT)扩增形成聚胸腺嘧啶(T)DNA模板,制备了聚T铜纳米簇(TS-CuNCs),构建了一种用于L-组氨酸(L-His)检测的荧光传感分析新方法。TdT酶在d TTP存在下合成聚T单链DNA核苷酸序列。由于胸腺嘧啶和Cu~(2+)之间的亲合力,聚T单链DNA作为合成铜纳米簇(CuNCs)的模板,加入还原剂后形成CuNCs,荧光强度增强。在L-His存在下,L-组氨酸的咪唑基与Cu~(2+)螯合形成L-His-Cu~(2+)配合物,因而进入聚胸腺嘧啶序列中的Cu~(2+)量减少,使得合成的CuNCs数量减少,导致荧光信号减弱。实验结果表明,体系荧光响应信号与L-His浓度的对数值在5.0×10~(-9)~5.0×10~(-4)mol/L范围内呈线性关系,检出限达到3.4×10~(-9)mol/L。本方法用于实际尿液样品中L-组氨酸检测的回收率为97.4%~104.6%,在生物医学及临床诊断中具有潜在应用价值。
[Abstract]:The polythymidine (PTT) DNA template was amplified by terminal deoxynucleotidyl transferase (TdT). A novel fluorescent sensing method for the detection of Lhistidine L-His1 was constructed. The nucleotide sequence of poly T single strand DNA was synthesized by TDT enzyme in the presence of d TTP. Due to the affinity between thymine and Cu~(2, a novel fluorescent sensing method for the detection of Lhistidine L-His1 was constructed. Poly (T) single-stranded DNA was used as template for the synthesis of copper nanoclusters (CuNCs2), and the reduction agent was added to form CuNCs.The fluorescence intensity was enhanced. In the presence of L-His, the imidazolyl of L-histidine chelated with Cu~(2) to form L-His-CuNCs2) complex. As a result, the amount of Cu~(2 entering the polythymidine sequence decreased, the number of synthesized CuNCs decreased and the fluorescence signal weakened. The experimental results showed that the logarithmic value of the fluorescence response signal and L-His concentration was linear in the range of 5.0 脳 10 ~ (-9) ~ 5 脳 10 ~ (10) ~ (-4) mol 路L ~ (-1). The detection limit was 3.4 脳 10 ~ (-9) mol 路L ~ (-1). The recoveries of L- histidine in urine samples were 97.4 ~ 104.6, which had potential application value in biomedical and clinical diagnosis.
【作者单位】：长沙理工大学化学与生物工程学院电力与交通材料保护湖南省重点实验室微纳生物传感与食品安全检测协同创新中心;中国科学院亚热带农业生态研究所;
【基金】：国家自然科学基金项目(Nos.31527803,21545010,21645009) 中国科学院科技服务网络STS计划(No.KFJ-SW-STS-173)项目资助~~
【分类号】：O657.3;R446

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