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    STUDIA BIOLOGIA - Ediţia nr.1 din 2019  
         
  Articol:   TRANSCRIPTIONAL ANALYSIS OF glnA DELETION MUTANT IN HALOFERAX MEDITERRANEI.

Autori:  VERÓNICA RODRÍGUEZ-HERRERO, VANESA BAUTISTA, MÓNICA CAMACHO, MÓNICA CORTÉS, MARÍA-JOSÉ BONETE, JULIA ESCLAPEZ. 		 
       
         
  Rezumat:   Haloferax mediterranei (ATCC 33500T) genome contains three genes which present sequence homology with glutamine synthetase: glnA, glnA2 and glnA3. Deletion mutants of glnA gene were constructed to determine the role of this enzyme in the nitrogen and amino acid metabolism. The mutant HM26-glnA has been characterised using different approaches, concluding that glnA is an essential gene to the growth of H. mediterranei. The transcriptomes of deletion mutant HM26-glnA and parental strain (HM26) have been compared in different growth conditions. Statistical analysis of the global gene expression was carried out obtaining different expression profiles depending on the contrast analysed, showing up and down-regulated genes. In addition, a functional gene analysis was performed to find out the processes in which they are involved. The microarray data was confirmed by quantitative real-time PCR. The transcriptomes comparison results of HM26-ΔglnA against HM26 in the presence of nitrogen source shows that 3 genes are up -regulated and 50 are down-regulated. The great majority of down-regulated genes are related to amino acid metabolism and the transport system. However, the up-regulated genes were related to sulphur metabolism. The analysis of the data in the nitrogen absence shows changes in the transcription level of great number of genes in both HM26-ΔglnA and HM26. In this condition, the mutant shows 91 up-regulated and 79 down-regulated exclusive genes, which are mostly related to nitrogen metabolism compounds, transporters and regulation processes. These results suggest that the HM26-ΔglnA uses different metabolic strategies from that of the parental strain under nitrogen starvation conditions.

Keywords: eletion mutant, glutamine synthetase, microarray, nitrogen metabolism. 		 
         
     
         
         
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