Jongsma Maarten A, Litjens Ralph H G M
Plant Research International, Wageningen, The Netherlands.
Proteomics. 2006 May;6(9):2650-5. doi: 10.1002/pmic.200500654.
The high-throughput deposition of recombinant proteins on chips, beads or biosensor devices would be greatly facilitated by the implementation of self-assembly concepts. DNA-directed immobilization via conjugation of proteins to an oligonucleotide would be preeminently suited for this purpose. Here, we present a unique method to attach a single DNA address to proteins in one step during the purification from the E. coli lysate by fusion to human O6-alkylguanine-DNA-alkyltransferase (SNAP-tag) and the Avitag. Use of the conjugates in converting a DNA chip into a protein chip by self assembly is demonstrated.
自组装概念的应用将极大地促进重组蛋白在芯片、珠子或生物传感器装置上的高通量沉积。通过蛋白质与寡核苷酸的缀合进行DNA定向固定将非常适合此目的。在此,我们提出了一种独特的方法,在从大肠杆菌裂解物中纯化过程中,通过与人O6-烷基鸟嘌呤-DNA烷基转移酶(SNAP标签)和Avi标签融合,一步将单个DNA地址连接到蛋白质上。证明了通过自组装将DNA芯片转化为蛋白质芯片时这些缀合物的用途。