Generation of a recombinant Aggregatibacter actinomycetemcomitans RTX toxin in Escherichia coli.

ABSTRACT: A leukotoxin (LtxA) that is produced by Aggregatibacter actinomycetemcomitans (Aa) is an important virulence determinant in an aggressive form of periodontitis in adolescents. Understanding the function of this protein at the molecular level is critical to elucidating its role in the disease process. To accomplish genetic analysis of the protein structure and relating these observations to toxin function, we have developed an E. coli expression system for the generation and rapid purification of LtxA. Cloning the structural toxin gene, ltxA, from Aa strain JP2 under control of T7 promoter-1 of pCDFDuet-1 vector resulted in expression of a 114 KDa protein which could be easily purified by the presence of a carboxy-terminal engineered double hexahistidine (double-His₆) tag and was

SUBMITTER: Balashova N 

PROVIDER: S-EPMC6640136 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature