Borén T, Elias P, Samuelsson T, Claesson C, Barciszewska M, Gehrke C W, Kuo K C, Lustig F
Department of Medical Biochemistry, University of Göteborg, Sweden.
J Mol Biol. 1993 Apr 5;230(3):739-49. doi: 10.1006/jmbi.1993.1196.
To investigate the reading properties of adenosine in the wobble position we have used site-directed mutagenesis of the Escherichia coli glycine tRNA1(CCC) gene to substitute the nucleotide A in the wobble position of the corresponding tRNA. The effect of this change on the ability of the tRNA to discriminate between the nucleotides in the third position of the glycine codons has been investigated. We have compared the ability of the mutant glycine tRNA1(UCC) and glycine tRNA1(ACC) as well as the mycoplasma glycine tRNA(UCC) to read the glycine codons. The results showed that glycine tRNA1(ACC) unlike glycine tRNA1(UCC) did not fully discriminate between the glycine codons. These experiments were carried out using a new in vitro protein synthesizing system that allows us to monitor the reading of all four glycine codons. In the present paper we give a detailed description of this new in vitro system.
为了研究摆动位置上腺苷的阅读特性，我们利用定点诱变技术对大肠杆菌甘氨酸tRNA1(CCC)基因进行诱变，以替换相应tRNA摆动位置上的核苷酸A。研究了这种变化对tRNA区分甘氨酸密码子第三位核苷酸能力的影响。我们比较了突变型甘氨酸tRNA1(UCC)、甘氨酸tRNA1(ACC)以及支原体甘氨酸tRNA(UCC)阅读甘氨酸密码子的能力。结果表明，与甘氨酸tRNA1(UCC)不同，甘氨酸tRNA1(ACC)不能完全区分甘氨酸密码子。这些实验是使用一种新的体外蛋白质合成系统进行的，该系统使我们能够监测所有四个甘氨酸密码子的阅读情况。在本文中，我们对这种新的体外系统进行了详细描述。
