链霉亲和素磁珠的合成及其用于AMP适配子传感器的研究  

The Synthesis of Streptavidin Coated MBs and the Application on the Study of AMP Aptasensor

作　　者：张娟[1] 刘芳[1] 谌蓉[1] 邓乐[1] 

机构地区：[1]湖南师范大学生命科学学院,中国湖南长沙410081

出　　处：《生命科学研究》2010年第6期507-515,共9页Life Science Research

基　　金：国家自然科学基金资助项目(30770570);国家863项目(2007AA062Z403)

摘　　要：γ-Fe2O3是一种磁性极高的磁氧铁,被广泛用于磁性分离,然而将γ-Fe2O3磁性纳米粒子用于适配子生物传感器来研究微生物细胞中的低相对分子质量产物却鲜有报道.经非水相法合成的γ-Fe2O3磁性粒子成晶效果好,粒径为19 nm,具有良好的磁力.通过正硅酸乙酯:TEOS(ethyl silicate;tetraethyl orthosilicate)进一步处理形成在水相中分散好、粒径均匀、磁性优良的核壳型磁性纳米微球.修饰上链霉亲和素纳米微球与生物素修饰的DNA连接起来,可用于探讨和研究微生物体内的底物AMP(Adenosine Monophosphate).基于适配子与底物结合发生构象转变的原理对该适配子传感器灵敏度、特异性及活体细胞的研究进行了探讨,实验结果表明:这种新型的AMP适配子生物传感器的检测下限达到了纳摩级,且具有非常好的底物特异性,在活体中的检测亦呈现一定趋势.γ-Fe2O3 is one of the best magnetic materials.As a biomagnetic separation tool,it becoming increasingly important to be used in possible applications in the biosciences.However,this wonderful separation tool,together with the biosensors,used in the study on low-molecular-weight substrate in microorganisms,which is rarely reported.The 19 nm diameter nanoparticles of γ-Fe2O3,which are synthesized by non-aqueous phase,possessing good magnetic force.After treating by TEOS,particles would shape into magnetic beads（MBs）.If they are modified by streptavidin,these uniform grading and well dispersed MBs could have tight junction with biotin modified DNA,and they can be one of the important part of the biosensor to explore the AMP in microorganisms.As the constellation will transform to the special chair-style when aptamer binds to its ligands.Based on this character,the sensitive,specificity sensor,which is discussed and made use of in the analysis of AMP in microorganisms.The result showed that this AMP aptasensor possess good specificity,the detection limit come to nmol and the data in vivo also displayed good trend.

关 键 词：磁性纳米微球 适配子生物反应器 AMP 检测 

分 类 号：Q939.99[生物学—微生物学]