本文關(guān)鍵詞： 末端脫氧核苷酸轉(zhuǎn)移酶 聚胸腺嘧啶 銅納米簇 L-組氨酸 熒光分析　出處：《分析化學(xué)》2017年10期 　論文類型：期刊論文

【摘要】：利用末端脫氧核苷酸轉(zhuǎn)移酶(TdT)擴(kuò)增形成聚胸腺嘧啶(T)DNA模板,制備了聚T銅納米簇(TS-CuNCs),構(gòu)建了一種用于L-組氨酸(L-His)檢測(cè)的熒光傳感分析新方法。TdT酶在d TTP存在下合成聚T單鏈DNA核苷酸序列。由于胸腺嘧啶和Cu~(2+)之間的親合力,聚T單鏈DNA作為合成銅納米簇(CuNCs)的模板,加入還原劑后形成CuNCs,熒光強(qiáng)度增強(qiáng)。在L-His存在下,L-組氨酸的咪唑基與Cu~(2+)螯合形成L-His-Cu~(2+)配合物,因而進(jìn)入聚胸腺嘧啶序列中的Cu~(2+)量減少,使得合成的CuNCs數(shù)量減少,導(dǎo)致熒光信號(hào)減弱。實(shí)驗(yàn)結(jié)果表明,體系熒光響應(yīng)信號(hào)與L-His濃度的對(duì)數(shù)值在5.0×10~(-9)~5.0×10~(-4)mol/L范圍內(nèi)呈線性關(guān)系,檢出限達(dá)到3.4×10~(-9)mol/L。本方法用于實(shí)際尿液樣品中L-組氨酸檢測(cè)的回收率為97.4%~104.6%,在生物醫(yī)學(xué)及臨床診斷中具有潛在應(yīng)用價(jià)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.
【作者單位】： 長(zhǎng)沙理工大學(xué)化學(xué)與生物工程學(xué)院電力與交通材料保護(hù)湖南省重點(diǎn)實(shí)驗(yàn)室微納生物傳感與食品安全檢測(cè)協(xié)同創(chuàng)新中心;中國(guó)科學(xué)院亞熱帶農(nóng)業(yè)生態(tài)研究所;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(Nos.31527803,21545010,21645009) 中國(guó)科學(xué)院科技服務(wù)網(wǎng)絡(luò)STS計(jì)劃(No.KFJ-SW-STS-173)項(xiàng)目資助~~
【分類號(hào)】：O657.3;R446

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