首页> 美国卫生研究院文献>Acta Crystallographica Section F: Structural Biology and Crystallization Communications >Purification and crystallization of a putative transcriptional regulator of the benzoate oxidation pathway in Burkholderia xenovorans LB400
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Purification and crystallization of a putative transcriptional regulator of the benzoate oxidation pathway in Burkholderia xenovorans LB400

机译:异种伯克霍尔德氏菌LB400中苯甲酸酯氧化途径的推定转录调节子的纯化和结晶

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摘要

Burkholderia xenovorans LB400 harbours two paralogous copies of the recently discovered benzoate oxidation (box) pathway. While both copies are functional, the paralogues are differentially regulated and flanked by putative transcriptional regulators from distinct families. The putative LysR-type transcriptional regulator (LTTR) adjacent to the megaplasmid-encoded box enzymes, Bxe_C0898, has been produced recombinantly in Escherichia coli and purified to homogeneity. Gel-filtration studies show that Bxe_C0898 is a tetramer in solution, consistent with previously characterized LTTRs. Bxe_C0898 crystallized with four molecules in the asymmetric unit of the P43212/P41212 unit cell with a solvent content of 61.19%, as indicated by processing of the X-ray diffraction data. DNA-protection assays are currently under way in order to identify potential operator regions for this LTTR and to define its role in regulation of the box pathway.
机译:伯克霍尔德菌xenovorans LB400具有最近发现的苯甲酸酯氧化(盒)途径的两个旁系同源拷贝。虽然两个副本都起作用,但是旁系同源物受到不同家族的推定转录调节因子的差异调节和侧翼。与大质粒编码的框酶Bxe_C0898相邻的推定LysR型转录调节子(LTTR)已在大肠杆菌中重组生产并纯化至同质。凝胶过滤研究表明,Bxe_C0898是溶液中的四聚体,与先前表征的LTTR一致。如X射线衍射数据处理所示,Bxe_C0898在P43212 / P41212晶胞的不对称单元中被四个分子结晶,溶剂含量为61.19%。目前正在进行DNA保护测定,以鉴定该LTTR的潜在操纵基因区域,并确定其在盒途径调控中的作用。

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