Wung C H, Hsu Y H, Liou D Y, Huang W C, Lin N S, Chang B Y
J Gen Virol. 1999 May;80 ( Pt 5):1119-1126. doi: 10.1099/0022-1317-80-5-1119.
The triple gene block protein 1 (TGBp1) encoded by open reading frame 2 of bamboo mosaic potexvirus (BaMV) was overexpressed in Escherichia coli and purified in order to test its RNA-binding activity. UV crosslinking assays revealed that the RNA-binding activity was present mainly in the soluble fraction of the refolded TGBp1. The binding activity was nonspecific and salt concentration-dependent: activity was present at 0-50 mM NaCl but was almost abolished at 200 mM. The RNA-binding domain was located by deletion mutagenesis to the N-terminal 3-24 amino acids of TGBp1. Sequence alignment analysis of the N-terminal 25 amino acids of the TGBp1 homologues of potexviruses identified three arginine residues. Arg-to-Ala substitution at any one of the three arginines eliminated most of the RNA-binding activity, indicating that they were all critical to the RNA-binding activity of the TGBp1 of BaMV.
竹花叶病毒（BaMV）开放阅读框2编码的三基因块蛋白1（TGBp1）在大肠杆菌中过表达并纯化，以测试其RNA结合活性。紫外线交联分析表明，RNA结合活性主要存在于重折叠的TGBp1的可溶部分。结合活性是非特异性的且依赖盐浓度：在0-50 mM NaCl时存在活性，但在200 mM时几乎完全丧失。通过缺失诱变将RNA结合结构域定位到TGBp1的N端3-24个氨基酸。对马铃薯X病毒属病毒TGBp1同源物的N端25个氨基酸进行序列比对分析，鉴定出三个精氨酸残基。三个精氨酸中的任何一个被精氨酸到丙氨酸的取代消除了大部分RNA结合活性，表明它们对BaMV的TGBp1的RNA结合活性都至关重要。
