Cloning and nucleotide sequencing of the secA gene from coryneform bacteria

Genetic analysis, techniques and applications Pub Date : 1999-03-01 DOI:10.1016/S1050-3862(98)00010-2

Miki Kobayashi , Nobutake Fugono , Yoko Asai , Hideaki Yukawa

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引用次数: 6

Abstract

Taking advantage of highly conserved domains present in the secA gene from Escherichia coli and Bacillus subtilis, we designed degenerate oligonucleotides (oligos) corresponding to these regions. These oligos were used as primers in PCR in order to amplify DNA sequences from Brevibacterium flavum MJ233 chromosomal DNA. The PCR product was used as a probe to recover genomic fragments from a λ library of Br. flavum MJ233. The complete nucleotide sequence (nt) of the cloned 5.3-kb EcoRI fragment containing the secA homolog from Br. flavum MJ233 indicated that the deduced gene product of the Br. flavum secA homolog is composed of 845 amino acids (aa) with a deduced molecular weight (MW) of 95 429. Comparison of this aa sequence to the corresponding sequences from E. coli and B. subtilis revealed a high degree of conservation and suggested that the Br. flavum secA homolog has putative ATP binding regions.

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棒状细菌secA基因的克隆及核苷酸序列分析

利用大肠杆菌和枯草芽孢杆菌secA基因中存在的高度保守结构域，我们设计了与这些区域对应的简并寡核苷酸(oligos)。利用这些寡核苷酸作为引物扩增黄短杆菌MJ233染色体DNA序列。PCR产物作为探针从Br λ文库中恢复基因组片段。flavum MJ233。克隆的含有Br的secA同源物的5.3 kb EcoRI片段的完整核苷酸序列(nt)。黄酮类化合物MJ233表明该基因的推断产物。黄酮(flavum secA)同源物由845个氨基酸(aa)组成，分子量(MW)为95 429。将该aa序列与大肠杆菌和枯草芽孢杆菌的相应序列进行比较，发现其具有高度的保守性。黄酮a同源物具有假定的ATP结合区。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Genetic analysis, techniques and applications

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