Kukolka Florian, Niemeyer Christof M
Universitat Dortmund, Fachbereich Chemie Biologisch-Chemische Mikrostrukturtechnik, Otto-Hahn-Str. 6, D-44227 Dortmund, Germany.
Org Biomol Chem. 2004 Aug 7;2(15):2203-6. doi: 10.1039/b406492e. Epub 2004 Jul 8.
A novel species of DNA--protein conjugate was synthesized by chemically linking DNA oligonucleotides to Aequorea victoria green fluorescent protein mutant EYFP. An additional cysteine was added to the C-terminus of the EYFP by genetic engineering and used to covalently attach amino-modified oligonucleotide with the aid of the heterobifunctional crosslinker sSMCC. EYFP maintained its fluorescence upon conjugation. The oligonucleotide provides an additional binding site to the fluorescent protein, and hence, the EYFP conjugate could be specifically hybridized with both complementary DNA-protein conjugates in-solution as well as immobilized at capture oligonucleotides attached to a solid substrate. These studies are paving the way for future applications in the self-assembly of photoactive supramolecular complexes, such as artificial light-harvesting systems.
通过将DNA寡核苷酸与维多利亚水母绿色荧光蛋白突变体EYFP进行化学连接，合成了一种新型的DNA-蛋白质共轭物。通过基因工程在EYFP的C末端添加了一个额外的半胱氨酸，并借助异双功能交联剂sSMCC将氨基修饰的寡核苷酸共价连接。共轭后，EYFP保持其荧光。寡核苷酸为荧光蛋白提供了一个额外的结合位点，因此，EYFP共轭物可以与溶液中的互补DNA-蛋白质共轭物特异性杂交，也可以固定在附着于固体基质的捕获寡核苷酸上。这些研究为光活性超分子复合物（如人工光捕获系统）的自组装的未来应用铺平了道路。
