Genetic Variability of Zymomonas mobilis UFPEDA strains by 16S rDNA Gene Sequencing

Abstract

Traditionally, several phenotypic techniques have been applied to investigate bacterial characterization, however, advances in molecular techniques have enabled higher levels of specificity and reliability. In this case, most methods used include the application of PCR (Polymerase Chain Reaction) studying genetic-molecular markers, among them stands the 16S rDNA, because of its high intraspecific conservation. The bacterium Z. mobilis has attracted considerable interest due to its biotechnology potential. The genetic variability among seven strains of Zymomonas mobilis UFPEDA was analyzed by the sequencing and the restriction patterns of 16S rDNA. Each strain was grown in SSDL for 24 hours at 30 degrees, followed by centrifugation and DNA extraction. PCR reactions were performed with primers and conditions for amplification of the 16S rDNA. The fragments were sequenced and analyzed with the programs BLASTn, MultAlin. The theoretical restriction patterns of restriction endonucleases HaeIII, NdeII and StuI were generated from DistinctiEnz. Finally, a cladogram was built using the program ClustalW by neighbor-joining method. It was verified the degree of conservation of the 16S rDNA of strains of Z. mobilis, and can be observed several events of mutations that certainly was responsible for the different pattern of cleavage between the UFPEDA lines o and the lines of European collections, it is not possible subtyping the first by this technique. This work evidences genetic differences in UFPEDA strains which to be related to the physiological pattern aiming to improve the biotechnology ability of this collection.