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Etude de l'endoribonucléase de restriction RegB.

Abstract : The restriction endoribonuclease RegB from the bacteriophage T4 is involved in the transition between the early and the middle phase of the lytic cycle of the virus. RegB cleaves with an almost absolute specificity the GGAG sequence needed for the bacterial initiation of the translation. We characterized the toxicity level of RegB in the bacterium and presented new tools to bypass this problem. We also presented a biotechnological application of RegB for the design of a positive selection and dual expression vector. Using 31P NMR, we found that RegB acts as a transphosphorylase. Mutagenesis studies combined with the sequencing of the regB gene from the phage RB49 showed that the residues E19, H48 , R52 and H68 may be involved in the activity of RegB. The RegB H48A point mutant was chosen to build a structural model of the active site of RegB. The sequential assignment of the protein using 1H/15N/13C heteronuclear NMR was driven successfully.
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Submitted on : Wednesday, July 21, 2010 - 9:07:54 AM
Last modification on : Thursday, December 2, 2021 - 2:26:03 PM
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  • HAL Id : pastel-00000707, version 1


Fakhri Saïda. Etude de l'endoribonucléase de restriction RegB.. Sciences du Vivant [q-bio]. Ecole Polytechnique X, 2003. Français. ⟨pastel-00000707⟩



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