Analysis of translation mechanism for selenoprotein synthesis

我们成功地制备了选择性半胱氨酸掺入核糖体所需的蛋白质和RNA分子。这些分子是SBP2、L30、EFSec、SEICIS-RNA、tRNASec，并进行了生化和生物物理实验。可以分析和发表L30的晶体结构。由于我们最初成功地获得了具有功能的tRNASec，因此在特定氨基酸存在的情况下，tRNASec、EFSec和SBP2之间的结合实验正在进行中。用ITC观察到野生型与GDP有显著的结合亲和力(K_d=229 nM)，从而确定了制备该分子的方法。此外，为结晶目的而产生的突变体被证明具有适当的二级结构。

We successfully prepared protein and RNA molecules which are required for selecocystein incorporation on ribosome. These molecules are SBP2, L30, EFSec, SEICIS-RNA, tRNASec and subjected to biochemical and biophysical experiments. Crystal structure of L30 could be analyzed and published. Since we originally succeeded generating functional tRNASec, binding assay among tRNASec, EFSec, and SBP2 in the presence of specific amino acid are now in progress. We observed the significant binding affinity(K_d=229 nM) between the wild type and GDP using ITC, the way to prepare the molecule was thus confirmed. Furthermore, mutants generated for the purpose of crystallization were proved that they have proper secondary structures