Purpose We have previously shown that DNA constructs can be introduced into isolated mitochondria through the process of conjugative transfer from an E. coli host. We set out to generate a conjugative E. coli strain that would be able to introduce itself into the cytoplasm of a mammalian cell for the purpose of transferring DNA into the mitochondria in the cell. Methods We have now developed a method for making E. coli strains from which nonreplicating populations of daughter cells can be generated. We used this approach to modify a facultative intracellular enteroinvasive E. coli (EIEC) and introduced conjugative functions to this new strain. Results We demonstrate that this new strain can generate large populations of nonreplicating cells that are capable of conjugative transfer to other cells and can readily invade mammalian tissue culture cells, live in the cytoplasm of the cell for several days, and that do not kill the invaded mammalian cell. Conclusions We successfully constructed an E. coli host suitable for intracellular conjugative transfer but, due to the lack of suitable mitochondrial screening or selectable markers, we have not yet been able to determine if these bacterial vectors can in fact transfer DNA into intracelluar mitochondria.
Endothelial nitric oxide synthase polymorphism G298T in association with oxidative DNA damage in coronary atherosclerosis
Content Type Journal Article
Category Research Note
RAJESH G. KUMAR, Department of Genetics, Osmania Univers ... more
A micropropagation protocol was developed for multiplication of seedless lemon (Citrus limon L. cv. Kaghzi Kalan) using nodal explants. The maximum shoot regeneration was observed on low level of BAP (0.1 mg l−1) or kinetin (0.5 mg l−1). BA was recorded to be better than kinetin in terms of ... more
The objective of this study was to evaluate the activity, kinetic behavior, and thermal inactivation kinetics of peroxidase (POD) in aqueous extracts from two kinds of milk ripe stage corn, sweet corn and waxy corn. Optimum activities using guaiacol as the hydrogen donor were obtained for s ... more