Furthermore the distribution of natural competence is surprisingly sporadic; most naturally competent bacteria have many relatives, including other strains of the same species, that cannot be transformed under laboratory conditions (for examples see [ 3- 6]).
The uptake specificity of some naturally competent bacteria can also guide inferences about the evolution of competence.
Competent bacteria can take up extracellular DNA fragments present in several environments during natural transformation.
Most DNA taken up by competent bacteria will also be used for nutrition after further degradation.
This potential to use DNA as a source of both nutrients and genetic novelty has important implications for the ecology and evolution of competent bacteria.
All PCR products were cloned into T overhang vectors, transformed into competent bacteria, with positives colonies identified by colony PCR and multiple clones per PCR product sequenced.
It would be interesting to determine the whether the RecA proteins from other naturally competent bacteria also behave like RecANg in vitro.
Competent bacteria were generated, as described above, with pelleted cells re-suspended using 10 mL ice-cold calcium chloride (0.1 M).
Subsequently, the extracted plasmid mixture was transformed into DH5α competent bacteria and equally spread on cm (chloramphenical) and kan (kanamycin) containing LB agar plates.
After DNPI-mediated degradation of the plasmid template, we circularized the phosphorilated PCR product by self-ligation and used it to transform competent bacteria.
Finally the PCR product was treated with 1 µl DpnI endonuclease for 2 h at 37°C, and 2 µl DpnI-digested DNA was transformed into 50 µl DH5α competent bacteria.
