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嗜热栖热菌YT-1的recA基因：克隆、表达及特性分析

The recA gene from the thermophile Thermus aquaticus YT-1: cloning, expression, and characterization.

作者信息

Angov E, Camerini-Otero R D

机构信息

Genetics and Biochemistry Branch, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, Maryland 20892.

出版信息

J Bacteriol. 1994 Mar;176(5):1405-12. doi: 10.1128/jb.176.5.1405-1412.1994.

DOI:10.1128/jb.176.5.1405-1412.1994
PMID:8113181
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC205206/
Abstract

We have cloned, expressed, and purified the RecA analog from the thermophilic eubacterium Thermus aquaticus YT-1. Analysis of the deduced amino acid sequence indicates that the T. aquaticus RecA is structurally similar to the Escherichia coli RecA and suggests that RecA-like function has been conserved in thermophilic organisms. Preliminary biochemical analysis indicates that the protein has an ATP-dependent single-stranded DNA binding activity and can pair and carry out strand exchange to form a heteroduplex DNA under reaction conditions previously described for E. coli RecA, but at 55 to 65 degrees C. Further characterization of a thermophilically derived RecA protein should yield important information concerning DNA-protein interactions at high temperatures. In addition, a thermostable RecA protein may have some general applicability in stabilizing DNA-protein interactions in reactions which occur at high temperatures by increasing the specificity (stringency) of annealing reactions.

摘要

我们已经从嗜热真细菌嗜热水生栖热菌YT-1中克隆、表达并纯化了RecA类似物。对推导的氨基酸序列分析表明，嗜热水生栖热菌的RecA在结构上与大肠杆菌的RecA相似，这表明RecA样功能在嗜热生物中得以保留。初步生化分析表明，该蛋白具有依赖ATP的单链DNA结合活性，并且在先前描述的大肠杆菌RecA的反应条件下，但在55至65摄氏度时，能够配对并进行链交换以形成异源双链DNA。对嗜热来源的RecA蛋白的进一步表征应能产生有关高温下DNA-蛋白质相互作用的重要信息。此外，一种热稳定的RecA蛋白可能在通过提高退火反应的特异性（严格性）来稳定高温下发生的反应中的DNA-蛋白质相互作用方面具有一些普遍适用性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f33e/205206/43b0d32d60d3/jbacter00023-0216-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f33e/205206/6641e43c6f09/jbacter00023-0215-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f33e/205206/43b0d32d60d3/jbacter00023-0216-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f33e/205206/6641e43c6f09/jbacter00023-0215-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f33e/205206/43b0d32d60d3/jbacter00023-0216-a.jpg

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