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空肠弯曲菌组蛋白样蛋白编码基因hup的克隆与表达

Cloning and expression of the hup encoding a histone-like protein of Campylobacter jejuni.

作者信息

Konkel M E, Marconi R T, Mead D J, Cieplak W

机构信息

Laboratory of Vectors and Pathogens, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Hamilton, MT 59840.

出版信息

Gene. 1994 Aug 19;146(1):83-6. doi: 10.1016/0378-1119(94)90837-0.

Abstract

A Campylobacter jejuni gene, designated hup, that appears to encode a homolog of the histone-like DNA-binding protein, HU, has been cloned, sequenced and expressed in Escherichia coli. Immunoblotting and in vitro transcription/translation analyses revealed a 11-kDa protein that was produced by recombinant plasmids containing hup. The gene contains an open reading frame (ORF) sufficient to encode a protein of 98 amino acids (aa) with a calculated molecular mass of 10,267 Da and a predicted isoelectric point of 10.1. The deduced aa sequence of the protein, designated HCj, exhibits considerable sequence identity with members of the HU family of proteins from other eubacterial species. The transcription start point was identified by primer extension analysis and appropriately spaced promoter sequences were found which exhibit considerable similarity to E. coli and Bacillus promoters. Southern hybridization analyses indicate that C. jejuni has a single copy of hup.

摘要

一种空肠弯曲菌基因,命名为hup,它似乎编码一种类组蛋白DNA结合蛋白HU的同源物,已被克隆、测序并在大肠杆菌中表达。免疫印迹和体外转录/翻译分析显示,含有hup的重组质粒产生了一种11 kDa的蛋白质。该基因包含一个开放阅读框(ORF),足以编码一个98个氨基酸(aa)的蛋白质,计算分子量为10267 Da,预测等电点为10.1。该蛋白质推导的氨基酸序列,命名为HCj,与其他真细菌物种的HU蛋白家族成员具有相当高的序列同一性。通过引物延伸分析确定了转录起始点,并发现了间隔适当的启动子序列,这些序列与大肠杆菌和芽孢杆菌的启动子具有相当高的相似性。Southern杂交分析表明,空肠弯曲菌有一个hup单拷贝。

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