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百日咳博德特氏菌毒力基因座编码蛋白的亚细胞定位及免疫检测

Subcellular localization and immunological detection of proteins encoded by the vir locus of Bordetella pertussis.

作者信息

Stibitz S, Yang M S

机构信息

Laboratory of Bacterial Toxins, Food and Drug Administration, Bethesda, Maryland 20892.

出版信息

J Bacteriol. 1991 Jul;173(14):4288-96. doi: 10.1128/jb.173.14.4288-4296.1991.

Abstract

The DNA sequence of the central regulatory locus vir of Bordetella pertussis predicts that three gene products, BvgA, BvgB, and BvgC, are encoded. Features of the predicted primary structures of these proteins and their homology to other two-component systems suggest that BvgA is located in the cytoplasm, BvgB is located in the periplasm, and BvgC spans the inner membrane. We have used gene fusions to the phoA and lacZ genes of Escherichia coli to investigate the subcellular localization and membrane topology of these proteins. PhoA fusion proteins were also purified and used to raise antibodies that allowed visualization of the vir-encoded polypeptides by Western immunoblotting. Our results have largely confirmed the predictions of the DNA sequence, with the exception that BvgB and BvgC were found to constitute one larger protein that was homologous to the sensor class of two-component systems. We propose that this protein be named BvgS (for sensor) and that its gene be named bvgS.

摘要

百日咳博德特氏菌中心调控位点vir的DNA序列预测其编码三种基因产物,即BvgA、BvgB和BvgC。这些蛋白质预测的一级结构特征及其与其他双组分系统的同源性表明,BvgA位于细胞质中,BvgB位于周质中,BvgC跨内膜。我们利用与大肠杆菌的phoA和lacZ基因的基因融合来研究这些蛋白质的亚细胞定位和膜拓扑结构。PhoA融合蛋白也被纯化,并用于制备抗体,通过Western免疫印迹法可对vir编码的多肽进行可视化。我们的结果在很大程度上证实了DNA序列的预测,但发现BvgB和BvgC构成了一种与双组分系统传感器类同源的更大蛋白质这一情况除外。我们建议将该蛋白质命名为BvgS(代表传感器),其基因命名为bvgS。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5fc3/208088/253ccf4ff7ad/jbacter00104-0057-a.jpg

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