枯草芽孢杆菌中硒代半胱氨酸tRNA提取方法的研究。

J Matsugi, K Murao
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引用次数: 2

摘要

在枯草芽孢杆菌中,硒氨酸半胱氨酸tRNA迄今尚未在全基因组序列中被鉴定出来(1)。为了探索硒氨酸半胱氨酸在枯草芽孢杆菌中的掺入系统,我们通过aa-tRNA特异性生物素化(2)和RT-PCR(3)相结合的方法筛选丝氨酸可接受的tRNA,找到一个未知的硒氨酸半胱氨酸tRNA。从丝氨酸可接受tRNA库中获得的cdna被扩增并克隆到质粒中,以读其序列。该方法得到了相应的已知丝氨酸trna cDNA文库,但未发现硒代半胱氨酸的候选基因。因此,这一结果与之前的数据(4)可能表明枯草芽孢杆菌中不存在硒代半胱氨酸tRNA,并且/或者硒的代谢与其他细菌中已知的硒代谢有很大不同。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
A study of the method to pick up a selenocysteine tRNA in Bacillus subtilis.

In Bacillus subtilis, selenocysteine tRNA has not been identified in a total genome sequence so far (1). To explore the system of selenocysteine incorporation in B. subtilis, we screened serine-acceptable tRNAs to find an unknown tRNA for selenocysteine by the combined method of specific biotinylation of aa-tRNA (2) and RT-PCR (3). cDNAs obtained from the serine-acceptable tRNA pool were amplified and cloned into plasmid to read its sequence. This procedure gave cDNA library corresponding known serine tRNAs, but no candidate for selenocysteine has been found. Thus, this result, together with the previous data (4), might reveal that there is no selenocysteine tRNA in B. subtilis and/or metabolism of selenium is considerably different from known one as seen in other bacteria.

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