Christensen J H, Hansen P K, Lillelund O, Thøgersen H C
Protein Engineering Research Centre, University of Aarhus, Denmark.
FEBS Lett. 1991 Apr 9;281(1-2):181-4. doi: 10.1016/0014-5793(91)80388-j.
An N-terminal fragment of Xenopus TFIIIA, containing domains 1-3, (TF 3), was expressed in E. coli. High yields of recombinant zinc finger protein was isolated, and its DNA binding activity for the internal control region (ICR) of the Xenopus 5S RNA gene, was demonstrated by band-shift experiments and DNase I footprinting analysis. TF 3 protects 20 bp of ICR against DNase I digestion. The limits of protection are from +77 to +96 on both coding and noncoding strand. This protection pattern is identical to the protection pattern obtained with TFIIIA in the overlapping region, showing that the 3-finger fragment accounts fully for the protein-DNA interactions in TFIIIA-5S RNA gene over this region.
非洲爪蟾TFIIIA的一个包含结构域1 - 3的N端片段（TF 3）在大肠杆菌中表达。分离得到了高产率的重组锌指蛋白，通过凝胶迁移实验和DNase I足迹分析证明了其对非洲爪蟾5S RNA基因内部控制区（ICR）的DNA结合活性。TF 3保护ICR的20 bp不被DNase I消化。保护范围在编码链和非编码链上均为从 +77到 +96。这种保护模式与在重叠区域用TFIIIA获得的保护模式相同，表明这个三指片段完全解释了该区域TFIIIA - 5S RNA基因中蛋白质 - DNA的相互作用。
