铅的极谱络合吸附波法检测DNA特定序列  

Detection for DNA Specific Sequences Using the Polarographic Complex Adsorptive Wave of Pb

作　　者：张波[1] 刘鹤[1] 焦奎[1] 付洵[1] 孙伟[1] 

机构地区：[1]青岛科技大学化学与分子工程学院,山东青岛266042

出　　处：《青岛科技大学学报（自然科学版）》2009年第2期95-100,共6页Journal of Qingdao University of Science and Technology:Natural Science Edition

基　　金：国家自然科学基金项目(20635020;20405008);教育部博士点基金项目(20060426001)

摘　　要：应用极谱络合吸附波方法检测了草丁膦乙酰转移酶基因(PAT基因)的DNA特定序列。用水热法合成表面具有自由羧基的硒化铅(PbSe)纳米粒子,以乙基-(3-二甲基丙基)碳二亚胺盐酸盐(EDC)为偶联活化剂,将其标记于合成的5′端氨基修饰的寡聚核苷酸片段上,制成DNA探针,用于检测互补的目标DNA序列。以十六烷基三甲基溴化铵(CTAB)阳离子表面活性剂修饰的碳糊电极作为基底电极,通过静电作用将来自于PAT基因的目标DNA特定序列固定在电极表面,并与标记有PbSe纳米粒子的互补DNA探针杂交。以铅-铜铁试剂-六次甲基四胺-HAc极谱络合吸附波测定氧化溶解Pb-Se得到的Pb2+,成功检测了PAT基因的DNA特定序列。测定目标DNA的线性范围为1.0×10-11至1.0×10-7mol.L-1,检测限为8.7×10-12mol.L-1(3σ)。此方法测定目标DNA序列检测限低,灵敏度高,线性范围宽,重现性好,操作简捷方便,对互补和非互补序列具有很好的识别能力。The polarographic complex adsorptive wave was firstly used to detect DNA specific sequences of the phosphinothricin acetyltransferase（PAT）gene. Lead selenide （PbSe） nanoparticle surface-modified by free carboxyl was hydrothermally synthesized, and used as a marker to label the DNA probe for identifying complementary target DNA sequence. Sequence-specific DNA related to the PAT gene was used as target DNA sequence and immobilized electrostatically on cetyltrimethylammonium bromide （CTAB） modified carbon paste electrode （CTAB/CPE）, and further hybridized with the complementary probe DNA labeled with PbSe nanoparticles. The hybridization events were monitored by the oxidation dissolution of lead selenide anchored on the hybrids and Pb- cupferron-methenamine-acetic acid （HAe） polarographic complex adsorptive wave. DNA specific sequences related to the PAT gene was determined with a detection range from 1.0×10^-11 to 1.0×10^-2 mol·L^-1, and a detection limit of 8.7×10^-12 mol·L^-1（3σ）. The proposed method showed a good distinguishable ability to the noneomplemen-tary DNA sequences with the complementary DNA sequences.

关 键 词：DNA检测 极谱络合吸附波 硒化铅纳米粒子 DNA杂交 PAT基因 

分 类 号：O657.1[理学—分析化学]