Sequencing the Genome of a Marine Bacterium Called Pseudoalteromonasulvae TC14 and Molecular Characterization of Its Communication System (Quorum Sensing)
The communication system known as quorum sensing (QS) in gram-negative bacteria regulates biofilm formation and many other functions. The particularity of Pseudoalteromonasulvae TC14 seems to be the absence of a LuxI inducer, and therefore the lack of production of small molecules of Acyl homoserine lactone (AHL). Previous studies had shown that it does not harbor the inducer of the regulatory molecule (AHL). Nevertheless, it is able to regulate these same functions (biofilm, violacein) via AHLs. This could mean the presence of specific receptors for these induction molecules, belonging to the LuxR family. The aim of this study was to test these hypotheses using molecular analysis. Genome sequencing of the P. ulvae TC14 strain was carried out by Molecular Research LP (MR DNA), using the Illumina Hiseq 2500 method. The results revealed the presence of 2,293,242 base pairs, i.e. 100% of the genomic volume. The number of coding gene sequences was 1983, and the Guanine + Cytosine (G+C) percentage of the base number was 41.55%, revealing stability in the sequenced genome. In order to verify the identity of the sequenced genome, a phylogenetic analysis based on RNA16S was carried out. This analysis resulted in 93% homology with the previously sequenced and characterized species Pseudoalteromonastunicata, showing that it belongs to the genus Pseudoalteromonas. To these comparative results should be added those derived from genome analysis based on nucleotide percentage using the tools available at https://img.jgi.doe.gov/. The results showed that Pseudoalteromonastunicata D2 has the highest percentage nucleotide identity (ANI) (75.7913%), followed by Pseudoalteromonasflavipulchra NCIMB2033 (72.2736%) and Pseudoalteromonasphenolica KCTC 12086 (71.6685%). Next, the search for the various genes involved in QS was carried out using sequence comparisons via the BLAST method. For LuxI, sequence templates from the genomes of Vibrio, Chromobacterium, Shewanella or even other Pseudoalteromonas species were screened against the TC14 genome. This yielded no conclusive results synonymous with the non-presence of LuxI in TC14 as assumed by previous research. The search for LuxR, on the other hand, gave rise to the presence of eight presumptive sequences. Molecular characterization of the presumptive LuxR sequences was carried out to assess their gene expression. This characterization showed
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