Corbi N, Perez M, Maione R, Passananti C
Dipartimento di Biotecnologie Cellulari ed Ematologia, Università di Roma La Sapienza, Rome, Italy.
FEBS Lett. 1997 Nov 3;417(1):71-4. doi: 10.1016/s0014-5793(97)01257-x.
Using two synthetic oligonucleotides, we have constructed a new gene containing three zinc finger motifs of the Cys2-His2 type. We named this artificial gene 'Mago'. The Mago nucleotide triplets encoding the amino acid positions, described to be crucial for DNA binding specificity, have been chosen on the basis of the proposed recognition 'code' that relates the zinc finger's primary structure to the DNA binding target. Here we demonstrate that Mago protein specifically binds the 'code' DNA target, with a dissociation constant (Kd) comparable to the Kd of the well known Zif268 protein with its binding site. Moreover, we show that the deduced Mago 'code' and the 'experimental' selected DNA binding sites are almost identical, differing only in two nucleotides at the side positions.
我们使用两条合成寡核苷酸构建了一个新基因，该基因包含三个Cys2-His2型锌指基序。我们将这个人工基因命名为“Mago”。编码氨基酸位置的Mago核苷酸三联体被认为对DNA结合特异性至关重要，这些三联体是根据将锌指一级结构与DNA结合靶点相关联的推测识别“密码”来选择的。在此，我们证明Mago蛋白特异性结合“密码”DNA靶点，其解离常数（Kd）与著名的Zif268蛋白与其结合位点的Kd相当。此外，我们表明推导的Mago“密码”与“实验”选择的DNA结合位点几乎相同，仅在侧翼位置的两个核苷酸上有所不同。
