HIV-1电化学活性-非活性开关分子信标的构建及验证

The construction and confirmation of HIV-1 electrochemically active-inactive switching molecular beacon

目的构建人类免疫缺陷病毒1型(HIV-1)电化学活性-非活性开关分子信标(CAs-MB)并验证其结构及功能。方法合成并纯化针对HIV-1 gag基因保守区序列的CAs-MB,用傅里叶变换红外光谱(FT-IR)和循环伏安法(CV)验证其结构,用差分脉冲伏安法(DPV)信号强度评价CAs-MB的识别功能。结果胭脂红酸(CA)单体与CAs-MB FT-IR图的变化证明CA已标记于MB末端,CV图证明CAs-MB电化学活性ON和OFF状态的存在。DPV结果证实其能够在电化学平台上实现对目标序列的特异性识别,且能够区分一个碱基的差异。结论 CAs-MB作为HIV-1 gag基因的特异性识别元件有望用于构建新型HIV-1电化学基因传感器。

Objective To construct human immunodeficiency virus-1 （HIV-1） electrochemically active-inactive switching molecular beacon （CAs-MB） and confirm its structure and function. Methods CAs-MB targeted conserved region of HIV-1 gag gene was prepared and purified. The structure was confirmed by Fourier transform-infrared spectroscopy （FF-IR） and cyclic voltmnmetry （CV）, while its function of recognizing targets was evaluated by differential pulse vohammetry （DPV）. Results The variations of FF-IR spectra of free carminicacid （CA） and CAs-MB indicated that CA had covalently linked onto the ends of MB. The results of CV proved that both ON and OFF statuses of CAs-MB were existing. DPV results illustrated that CAs-MB was effective in signaling the presence of complementary target and discriminating targets in electrochemical gene biosensor that differ by a single nucleotide. Conclusions CAs-MB opens a new detective technique and can be expanded to electrochemical gene biosensor to perform HIV-1 gag gene specificity detection.