(1. 湖南大學(xué) 環(huán)境科學(xué)與工程學(xué)院，長(zhǎng)沙 410082；2. 湖南城市學(xué)院 化學(xué)與環(huán)境工程學(xué)院，益陽(yáng) 413000；3. 湖南大學(xué) 環(huán)境生物與控制教育部重點(diǎn)實(shí)驗(yàn)室，長(zhǎng)沙 410082)
摘 要: 采用分子自組裝技術(shù)制備巰基修飾的基因分子膜修飾金電極，用于競(jìng)爭(zhēng)式雜交檢測(cè)黃孢原毛平革菌木素過(guò)氧化物酶編碼基因。通過(guò)差分脈沖伏安法、循環(huán)伏安法、交流阻抗法和電流—時(shí)間曲線法優(yōu)化自組裝時(shí)間和信號(hào)探針的最佳響應(yīng)濃度，研究目標(biāo)基因的線性檢測(cè)范圍和再生性能。結(jié)果表明：修飾金電極最優(yōu)自組裝時(shí)間為15 h，信號(hào)探針的最佳響應(yīng)濃度為0.51×10-6 mol/L，目標(biāo)基因的線性檢測(cè)范圍為7.51×10-12~1.05×10-9 mol/L，檢測(cè)下限為7.51×10-13 mol/L。該修飾電極具有良好的再生性能。
關(guān)鍵字: 修飾金電極；自組裝單分子膜；基因；競(jìng)爭(zhēng)雜交；電化學(xué)檢測(cè)
（1. College of Environmental Science and Engineering, Hunan University, Changsha 410082, China;2. College of Chemistry and Environmental Engineering, Hunan City University, Yiyang 413000, China;3. Key Laboratory of Environmental Biology and Pollution Control (Hunan University),Ministry of Education, Changsha 410082, China）
Abstract:A gold electrode with thiolated capture probe through self-assembling based on the competitive hybridization for detection of target sequence of lignin peroxidase (lip) gene of Phanerochaete chrysosporium was developed. Following hybridizations with competitively hybridized with the target nucleic acid and biotinylated response probe, streptavidin-horseradish peroxidase (HRP) conjugate was applied to the electrode. The electrochemical behavior was analyzed by cyclic voltammetry, electrochemical impedance spectroscopy, current—time curve and differential pulse voltammetry. The results show that the best time for self-assembling is 15 h, and the optimum concentration of response probe is 0.51×10-6 mol/L. A good linear correlation between the current and the concentration of the target nucleic acid concentration is found in the concentration range from 7.51×10-12 mol/L to 1.05×10-9 mol/L. The detection limit is 7.51×10-13 mol/L. The modified electrode exhibits high sensitivity, precision, stability and reproducibility.
Key words: modified gold electrode; self-assembled monolayer; gene; competitive hybridization; electrochemical determination
