Oswald T. Avery, an American pneumococcal researcher, speculated that Griffith’s experiment lacked appropriate control. However, subsequent, similar experiments carried out in Avery’s laboratory confirmed Griffith’s discovery. The experiments conducted later by Avery, MacLeod, and McCarty, and by Hershey and Chase proved that the transforming factor was DNA and elucidated its exact nature. Thereby, establishing the central role of DNA in inheritance.
When the mice were inoculated, the bacterial virulence was exhibited, causing pneumonia, and this eventually led to the death of the mice. On examining the blood of the deceased mice, progeny of the inoculated cells were obtained.
Type II R bacteria
When injected into the mice, the bacterial cells were successfully eliminated by the immune system, and hence, the mice lived. The blood showed no presence of the inoculated cells.
When the virulent strain was rendered avirulent by heating and killing it (heat-killed), and then injected into the mice, the strain did not show virulence, and was eliminated by the host’s immune system; hence, the mice survived. Their blood showed no presence of the inoculated cells.
Injecting the mice with a combination of equal number of cells of type II R strain and heat-killed type III S strain, caused pneumonia which progressed till the mice died. The bacterial cells isolated from the blood of these mice showed the presence of live type III S bacterial cells.
This mechanism differs slightly due to the difference in the structure of the cell membranes of the bacteria. Bacteria are broadly classified into two types based on this difference – Gram-negative and Gram-positive. The general outline is more or less similar. The presence of exogenous DNA is detected by the cell and natural competence is induced, then the foreign DNA binds to a DNA receptor on the surface of these competent cells. This receptor binding allows the activation of the DNA translocase system that allows the passing of DNA into the cell via the cell membrane. During this process, one strand of the DNA is degraded by the action of nucleases. The translocated single strand is then incorporated into the bacterial genome via the help of a RecA-dependent process.
Gram-negative bacteria show the presence of an extra membrane, hence, for DNA to be taken up, a channel is formed on the outer membrane by secretins. The uptake of a DNA fragment is generally not specific to its sequence; however, in some bacterial species, it has been seen that the presence of certain DNA sequences facilitate and enhance efficient uptake of the genetic material.
The divalent cations function to weaken the molecular structure of the cell membrane, hence, making it more permeable. The subsequent heat pulses cause the creation of a thermal imbalance, and in the process of regaining balance, the DNA molecules gain entry via the weakened membrane and into the cell.
Artificial competence can be alternatively induced and promoted via the use of a technique called electroporation. It involves applying an electric current to the cell suspension. This causes the formation of pores in the cell membrane. The exogenous DNA is taken up via these holes, which are resealed via the cell membrane repair machinery.
The process of electroporation can also be used for transformation purposes, and efficiency can be enhanced using enzymatic digestion or agitation using glass beads.
Plant cells can also be transformed using viral particles (transduction). Here, the genetic material to be inserted is packaged into a suitable plant virus. This modified virus is then allowed to infect the plant cells. The transfer occurs according to the viral machinery and transformation is achieved. Electroporation can also be used for plant cells.