Cloning of the trkAH gene cluster and characterization of the Trk K+-uptake system of Vibrio alginolyticus

Article Abstract:

Chromosomal DNA fragments of Vibrio alginolyticus were cloned into plasmids and the cells of an Escherichia coli strain defective in K+ uptake were electroporated and selected for growth at low K+ concentration to identify K+-uptake genes in V. alginolyticus. Results showed a 4.1 kb DNA fragment consisting of a cluster of three ORFs on the same DNA strand. It was also observed that the products of V. alginolyticus trkA and orf1 were present in E. coli minicells. The findings indicate that V. alginolyticus genes were functionally capable of replacing their E. coli counterparts.

author: Bakker, Evert P., Nakamura, Tatsunoke, Yamamuro, Naoya, Stumpe, Stefan, Unemoto, Tsutomu
Escherichia coli, Potassium channels, Vibrio, Chromosomal proteins

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Cloning of the Rhodobacter sphaeroides hisI gene: unifunctionality of the encoded protein and lack of linkage to other his genes

Article Abstract:

A nucleotide sequence analysis of the Rhodobacter sphaeroides 2.4.1 hisI gene found that it encodes a deduced product of 119 aa with a predicted molecular mass of 13*4 kDa. It was also found that the gene has no single histidine operon like that of Escherichia coli, encodes a unifunctional protein and is not linked to other his genes. The gene was isolated from a genomic library of R. sphaeroides through complementation of an Escherichia coli hisI mutant.

author: Barbe, Jordi, Oriol, Esther, Mendez-Alvarez, Sebastian, Gibert, Isidre
Chromosome mapping, Histidine

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Determination of the Paracoccus denitrificans SOS box

Article Abstract:

Gel retardation studies with crude extracts of Paracoccus denitrificans showed that a protein specifically binds to the promoter of the P. denitrificans recA gene. PCR mutagenesis of the recA promoter indicated that the GAACN7GAAC motif is needed for the formation of the DNA-protein complex. This protein also binds to the GTTCN7GTTC motif, which is found in the promoter of the P. denitrificans uvrA gene.

author: Barbe, Jordi, Rey, Alfonso del, Diestra, Julio, Fernandez de Henestrosa, Antonio R.
Genetic aspects, Bacteria, DNA damage

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subjects list: Research, Cloning
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