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深海细菌的高压生长需要RecD功能。

RecD function is required for high-pressure growth of a deep-sea bacterium.

作者信息

Bidle K A, Bartlett D H

机构信息

Marine Biology Research Division, Scripps Institution of Oceanography, University of California, San Diego, La Jolla, California 92093-0202, USA.

出版信息

J Bacteriol. 1999 Apr;181(8):2330-7. doi: 10.1128/JB.181.8.2330-2337.1999.

Abstract

A genomic library derived from the deep-sea bacterium Photobacterium profundum SS9 was conjugally delivered into a previously isolated pressure-sensitive SS9 mutant, designated EC1002 (E. Chi and D. H. Bartlett, J. Bacteriol. 175:7533-7540, 1993), and exconjugants were screened for the ability to grow at 280-atm hydrostatic pressure. Several clones were identified that had restored high-pressure growth. The complementing DNA was localized and in all cases found to possess strong homology to recD, a DNA recombination and repair gene. EC1002 was found to be deficient in plasmid stability, a phenotype also seen in Escherichia coli recD mutants. The defect in EC1002 was localized to a point mutation that created a stop codon within the recD gene. Two additional recD mutants were constructed by gene disruption and were both found to possess a pressure-sensitive growth phenotype, although the magnitude of the defect depended on the extent of 3' truncation of the recD coding sequence. Surprisingly, the introduction of the SS9 recD gene into an E. coli recD mutant had two dramatic effects. At high pressure, SS9 recD enabled growth in the E. coli mutant strain under conditions of plasmid antibiotic resistance selection and prevented cell filamentation. Both of these effects were recessive to wild-type E. coli recD. These results suggest that the SS9 recD gene plays an essential role in SS9 growth at high pressure and that it may be possible to identify additional aspects of RecD function through the characterization of this activity.

摘要

从深海细菌深渊发光杆菌SS9中获得的基因组文库通过接合转移至先前分离出的对压力敏感的SS9突变体(命名为EC1002,E. Chi和D. H. Bartlett,《细菌学杂志》175:7533 - 7540,1993年),并对接合后体筛选在280个大气压静水压力下生长的能力。鉴定出了几个恢复了高压生长能力的克隆。互补DNA被定位,并且在所有情况下都发现与recD(一个DNA重组和修复基因)具有很强的同源性。发现EC1002在质粒稳定性方面存在缺陷,这种表型在大肠杆菌recD突变体中也可见。EC1002中的缺陷定位到一个点突变,该突变在recD基因内产生了一个终止密码子。通过基因敲除构建了另外两个recD突变体,并且发现它们都具有压力敏感的生长表型,尽管缺陷的程度取决于recD编码序列3'端截短的程度。令人惊讶的是,将SS9 recD基因导入大肠杆菌recD突变体产生了两个显著的效果。在高压下,SS9 recD使大肠杆菌突变菌株在质粒抗生素抗性选择条件下能够生长,并防止细胞丝状化。这两种效果相对于野生型大肠杆菌recD都是隐性的。这些结果表明,SS9 recD基因在SS9高压生长中起重要作用,并且通过对这种活性的表征有可能确定RecD功能的其他方面。

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