Wei Yong, Ying Dajun, Hou Chunli, Zhu Chuhong, Cui Xiaoping, Xing Yan, Guo Hongfeng
Key Laboratory of Biomechanics and Tissue Engineering, Chongqing, China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2008 Jun;25(3):662-7.
This experiment was aimed to create A20 gene site-specific zinc finger DNA-binding protein. The sequence of A20 gene promoter was analyzed with bioinformatics means and submitted to ZF Tools Server at TSRI. Using the database of the web site, we determined the A20 gene valid target sites and designed the amino acid sequence of zinc finger protein predicted to be bound to the target site. And then, the structure of the protein sequence was analyzed and homology was modeled with various bioinformatics means. Based on the characteristic of this protein, the prokaryotic expression vector pTYB11-ZFP was constructed and expressed. Thus, the artificial zinc finger protein that recognized A20 specific sequence was designed, and expressed in Escherichia coli. The results indicate that it is feasible to design engineered artificial Zinc finger proteins by means of bioinformatics.
本实验旨在构建A20基因位点特异性锌指DNA结合蛋白。运用生物信息学方法分析A20基因启动子序列，并提交至TSRI的ZF Tools Server。利用该网站数据库，确定A20基因有效靶位点，并设计预测与靶位点结合的锌指蛋白氨基酸序列。然后，运用多种生物信息学方法分析该蛋白序列结构并进行同源建模。基于此蛋白特性，构建原核表达载体pTYB11-ZFP并进行表达。从而设计出识别A20特异性序列的人工锌指蛋白，并在大肠杆菌中表达。结果表明，通过生物信息学方法设计工程化人工锌指蛋白是可行的。
