Guild Katherine, Zhang Yang, Stacy Robin, Mundt Elizabeth, Benbow Sarah, Green Amanda, Myler Peter J
Seattle Biomedical Research Institute, USA.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Sep 1;67(Pt 9):1027-31. doi: 10.1107/S1744309111032143. Epub 2011 Aug 31.
Recombinant expression of proteins of interest in Escherichia coli is an important tool in the determination of protein structure. However, lack of expression and insolubility remain significant challenges to the expression and crystallization of these proteins. The SSGCID program uses a wheat germ cell-free expression system as a rescue pathway for proteins that are either not expressed or insoluble when produced in E. coli. Testing indicates that the system is a valuable tool for these protein targets. Further increases in solubility were obtained by the addition of the NVoy polymer reagent to the reaction mixture. These data indicate that this eukaryotic cell-free expression system has a high success rate and that the addition of specific reagents can increase the yield of soluble protein.
在大肠杆菌中重组表达目标蛋白是确定蛋白质结构的一项重要工具。然而，表达缺失和不溶性仍然是这些蛋白质表达和结晶的重大挑战。SSGCID程序使用小麦胚无细胞表达系统作为在大肠杆菌中不表达或不溶性蛋白质的拯救途径。测试表明该系统对于这些蛋白质靶点是一种有价值的工具。通过向反应混合物中添加NVoy聚合物试剂进一步提高了溶解度。这些数据表明这种真核无细胞表达系统具有很高的成功率，并且添加特定试剂可以提高可溶性蛋白质的产量。
