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Preparation crystallization and preliminary X-ray analysis of YjcG protein from Bacillus subtilis

机译:枯草芽孢杆菌YjcG蛋白的制备结晶和X射线初步分析

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摘要

Bacillus subtilis YjcG is a functionally uncharacterized protein with 171 residues that has no structural homologue in the Protein Data Bank. However, it shows sequence homology to bacterial and archaeal 2′–5′ RNA ligases. In order to identify its exact function via structural studies, the yjcG gene was amplified from B. subtilis genomic DNA and cloned into the expression vector pET21-DEST. The protein was expressed in a soluble form in Escherichia coli and was purified to homogeneity. Crystals suitable for X-ray analysis were obtained that diffracted to 2.3 Å and belonged to space group C2, with unit-cell parameters a = 99.66, b = 73.93, c = 61.77 Å, β = 113.56°.
机译:枯草芽孢杆菌YjcG是具有171个残基的功能上未表征的蛋白质,在蛋白质数据库中没有结构同源物。但是,它显示了与细菌和古细菌2'-5'RNA连接酶的序列同源性。为了通过结构研究确定其确切功能,从枯草芽孢杆菌基因组DNA中扩增了yjcG基因,并将其克隆到表达载体pET21-DEST中。蛋白质以可溶形式在大肠杆菌中表达,并纯化至均一。获得了适用于X射线分析的晶体,其衍射至2.3Å,属于C2空间群,其晶胞参数a = 99.66,b = 73.93,c = 61.77,β= 113.56°。

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