Bamdad C
Department of Biophysics, Harvard University, Cambridge, Massachusetts 02138, USA.
Biophys J. 1998 Oct;75(4):1997-2003. doi: 10.1016/S0006-3495(98)77641-6.
A novel method for DNA surface immobilization and a paradigm for the attachment of unmodified DNA of any length or sequence are described herein. The development of a DNA self-assembled monolayer (DNA-SAM) that incorporates a DNA-thiol into a monolayer of inert alkane thiolates is reported. This DNA-SAM specifically hybridized complementary oligonucleotides while resisting the nonspecific adsorption of noncomplementary DNA and irrelevant proteins. Duplex DNA, having a single-stranded "capture tail," specifically bound to the DNA-SAM if the sequence of the "tail" was complementary to DNA presented in the SAM. The sense strand of the hybridized duplex DNA could be covalently attached to the surface by an enzymatic ligation reaction (leaving the anti-sense strand dissociable). DNA-binding proteins specifically bound to these surfaces only if their cognate sites were present in the duplex DNA.
本文描述了一种用于DNA表面固定的新方法以及一种用于连接任何长度或序列的未修饰DNA的范例。报道了一种将DNA-硫醇掺入惰性烷硫醇单层中的DNA自组装单层（DNA-SAM）的开发。这种DNA-SAM能特异性杂交互补寡核苷酸，同时抵抗非互补DNA和无关蛋白质的非特异性吸附。具有单链“捕获尾”的双链DNA，如果“尾”的序列与SAM中呈现的DNA互补，则会特异性结合到DNA-SAM上。杂交双链DNA的有义链可通过酶促连接反应共价连接到表面（使反义链可解离）。只有当双链DNA中存在其同源位点时，DNA结合蛋白才会特异性结合到这些表面。
