单链脱氧核糖核酸探针在氧化锆陶瓷小珠表面的微阵列及陶瓷荧光小珠的制备  

Microarray of Single Stranded Deoxyribonuleic Acid Probes on the Surface of ZrO_2 Ceramic Beads and the Preparation of Fluorescent Ceramic Beads

作　　者：黄承志[1] 李原芳[1] 奉萍[1] 范美坤[1] 

机构地区：[1]西南师范大学环境化学研究所,重庆400715

出　　处：《分析化学》2002年第1期13-17,共5页Chinese Journal of Analytical Chemistry

基　　金：教育部高等学校骨干教师资助计划(文号:教技司2000-65);国家自然科学基金(No.20045002);重庆市科委应用基础

摘　　要：以Ｎ－（４－马来酰亚胺氧化丁酰）－琥珀酰亚胺磺酸钠盐（ｓｕｌｆｏ－ＧＭＢＳ）为偶联剂将５’－氨基／３’－得克萨斯红双末端修饰ＤＮＡ探针偶联到３－（２－氨乙基氨丙基）三甲氧基硅烷（ＡＡＴＳ）修饰的３００μｍＺｒＯ２陶瓷小珠表面，制成荧光小珠。小珠表面上单链ＤＮＡ探针分子之间的距离取决于小珠表面的预处理、偶联反应溶液中ＤＮＡ探针浓度、小珠大小和小珠的数目。在优化条件下，２０－ｍｅｒ单链ＤＮＡ探针在小珠表面微阵列的最小分子间距约为７．６ｎｍ，即在一个３００μｍＺｒＯ２陶瓷小珠表面有１０１０数量级的２０－ｍｅｒ单链 ＤＮＡ探针参与微阵列反应。机理研究表明：ＤＮＡ探针通过ｓｕｌｆｏＧＧＭＢＳ偶联到小珠表面的化学反应仅涉及到探针末端修饰氨基。The microarray features of 5'- NH2/3'- Texas red terminus modified deoxyribonucleic acid (DNA) probes on the surfaces of 300 mum ZrO2 ceramic beads were characterized through the coupling reaction of the probes with N-(4-maleimidobutyryloxy) sulfosuccininimide sodium salt (sulfa-GMBS). Based on the fluorescence measurement on the bead surface, it was found that the microarray of DNA probes depends on the pretreatment of the bead surface, DNA probe concentrations in aqueous solution and the total surface area of the beads. On optimal conditions, a 20-mer DNA probe can dot on the beads surface with the molecule distance of 7.6 nm, and more than 1.0 x 10(10) 20-mer single-stranded DNA probe can be microarrayed on one ZrO2 bead surface. Mechanism study shows that the coupling reaction of DNA probe on the ceramic bead surface only involves in the terminus NH2-group.

关 键 词：脱氧核糖核酸探针 陶瓷小珠 微阵列 DNA光谱探针 生物芯片 氧化锆陶瓷 

分 类 号：Q503[生物学—生物化学]