UAA编码氨基酸表达体系的构建

发布时间：2018-05-12 15:55

本文选题：UAA编码 + 表达体系　；参考：《吉林大学》2017年硕士论文

【摘要】：与所有的生命过程一样,外源基因在原核细胞中的表达包括两个主要过程,即DNA转录成mRNA和mRNA翻译成蛋白质。大肠杆菌一直是基因工程中应用最为广泛的表达系统。包含非编码氨基酸的蛋白质的过量表达已引起越来越多科学家的关注。非编码氨基酸的定点掺入,使关于蛋白质结构和功能的化学假定的特异检测成为可能。具有新化学性质的非编码氨基酸的定点掺入,可能使蛋白质产生新的功能。本文将pEVOL载体中对应琥珀密码子UAG的tRNA合酶进行定点突变,将其改造成对应赭石密码子UAA的tRNA/氨酰tRNA合酶表达系统,将UAA密码子变成了对应8-羟基喹啉丙氨酸的非编码氨基酸表达体系,并表达了掺入8-羟基喹啉丙氨酸的金属硫蛋白-GFP融合蛋白,验证了该蛋白的金属结合能力。
[Abstract]:As in all life processes, the expression of exogenous genes in prokaryotic cells involves two main processes: DNA transcription into mRNA and mRNA translation into proteins. E. coli has been the most widely used expression system in genetic engineering. Overexpression of proteins containing non-encoded amino acids has attracted more and more attention from scientists. The incorporation of non-coding amino acids makes possible the specific detection of chemical assumptions about the structure and function of proteins. The incorporation of non-coding amino acids with new chemical properties may produce new functions for proteins. In this paper, the tRNA synthase corresponding to the amber codon UAG in the pEVOL vector was mutated and transformed into the tRNA/ aminoyl tRNA synthase expression system corresponding to the ochre codon UAA. The UAA codon was transformed into a non-coding amino acid expression system corresponding to 8-hydroxyquinoline-alanine, and the fusion protein of metallothionein GFP was expressed with 8-hydroxyquinoline-alanine, which proved the metal-binding ability of the protein.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q78

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