期刊论文

Gong Ju-Fang

中文

生物化学与生物物理进展

1000-3282

2008

35

11

1320-1325

10.3321/j.issn:1000-3282.2008.11.015

Zhejiang Provincial Natural Science Foundation of China [Y204417]

本校为第一机构

为了研究tRNA^Trp的氨基酸接受茎中除两对半碱基以外的特异性元件，设计并完成了4种水稻线粒体tRNA^Trp向枯草杆菌tRNA^Trp的突变体（MPB0，G1A和U5G／A68C；MPB1，C2G／G71C；MPB2，C4G／G69C；MPB3，C2G／G71C和C4G／G69C），体外转录并用枯草杆菌和人这两种不同种属来源的色氨酰tRNA合成酶（TrpRS）N定了这些tRNA^Trp分子的氨酰化活力（Kcat／KM）．结果表明，这些突变体具有被枯草杆菌TrpRS氨酰化的能力，与野生型水稻线粒体tRNA^Trp相比，MPB0被枯草杆菌TrpRS氨酰化的活力提高了5倍，MPB1和MPB2被枯草杆菌TrpRS氨酰化的活力分别提高了40和53倍，MPB3则提高了140倍，为野生型枯草杆菌tRNA^T...

Four mutants from Oryza saliva mitochondria tRNA(TrP) toward B. subtilis tRNA(Trp) were constructed and transcribed in vitro with T7 RNA polymerase. The kinetic parameters (K-cat/K-M) of B. subtilis tryptophanyl-tRNA synthetase(TrpRS) and human TrpRS were determined with four mutant-type tRNA(Trps). Results showed that for reaction with B. subtilis TrpRS, C2/G71 and C4/G69 mutations each induced a comparable 40-fold and 53- fold of activity to Oryza saliva mitochondria tRNA(Trp) respectively. Notably, when the C2/G71 and C4/G69 mutations were introduced together into B. subtilis tRNATP, a 140-...