Site-specific uv crosslinking of minihelix DNA and TrpRS from Bacillus subtilis  

作　　者：Xu, F Jiang, G Zhang, Y Zhu, LF Jin, YX Wang, DB 

机构地区：[1]Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Shanghai 200031, Peoples R China

出　　处：《Chinese Science Bulletin》2001年第16期1380-1383,共4页

基　　金：This work was supported by the National Natural Science Foundation of China (Grant No. 39730120) ; the Chinese Academy of Sciences (Grant No. KSCX 2-2-04).

摘　　要：In order to investigate the recognition mechanism and the relationship between structure and function of minihelix DNA with Tryptophanyl-tRNA Synthetase (TrpRS), TrpRS from Bacillus Subtilis was purified. Four minihelix DNAs were chemically synthesized and the photoreactive reagent s4T was incorporated into three of them at the positions of G73, T72 and T55 corresponding to tRNATrp. The apparatus for uv crosslinking was devised and the parameters for uv crosslinking were optimized. The results indicated that the G73 and T72 base of minihelix DNA interacted with TrpRS directly. The uv crosslinking reaction was improved by the dose of uv irradiation and the concentration of both TrpRS and minihelix DNA.In order to investigate the recognition mechanism and the relationship between structure and function of minihelix DNA with Tryptophanyl-tRNA Synthetase (TrpRS), TrpRS from Bacillus Subtilis was purified. Four minihelix DNAs were chemically synthesized and the photoreactive reagent s4T was incorporated into three of them at the positions of G73, T72 and T55 corresponding to tRNATrp. The apparatus for uv crosslinking was devised and the parameters for uv crosslinking were optimized. The results indicated that the G73 and T72 base of minihelix DNA interacted with TrpRS directly. The uv crosslinking reaction was improved by the dose of uv irradiation and the concentration of both TrpRS and minihelix DNA.

关 键 词：UV CROSSLINKING minihelix DNA TrpRS discriminator. 

分 类 号：Q523[生物学—生物化学] Q503