Interaction between Repressor Protein ArgR and Arginine Biosynthesis Pathway Related Genes in Corynebacterium glutamicum ATCC 14067
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Abstract:
In bacteria, the repressor protein inhibits the transcription of arginine biosynthesis related genes through binding to conserved sequences in these genes. In Corynebacterium glutamicum ATCC 14067, the genes involved in arginine biosynthesis are located in argCJBDFRGH and carAB gene clusters. The recombinant E.coli BL21 (DE3)/ pET22b-argR was constructed to induce the expression of ArgR derived from ATCC 14067 and subsequently achieve the target protein. The analysis of the ArgR protein using molecular sieve columns showed that the relative molecular mass of ArgR protein was about 108 ku, and the ArgR protein was hexamer in the active state. The argC, argJ, argB, argF, argG, argH, carA gene fragments were obtained from biotin-labeled Corynebacterium glutamicum ATCC 14067, and their in vitro interactions with the ArgR protein were examined by electrophoretic mobility shift assays (EMSA). The results showed that the ArgR protein could bind to argC gene at positions -26 ~+32 bp, argB gene at positions -86~-29 bp, argG gene at positions -161~-104 bp, and carA gene at positions -99~ -42 bp, and the degree of binding to the argB gene was the greatest whilst no binding between ArgR and the argJ, argF or argH gene fragment. For the first time, the target binding sites of ArgR protein in argC, argB, argG, and carA genes were located within 60 bp, which provided a theoretical basis for further study on the regulatory mechanism of arginine synthesis in Corynebacterium glutamicum ATCC 14067.
