Firstly, the donor organism,’s DNA is cut by a restriction enzyme. The cut-up section is called “sticky-ends”, which enables the cut sections to meet up with other DNA strands (Mohan, 2005). Proceeding on, plasmids that induce tumour is treated with the same restriction enzyme, opening out the circle of DNA leaving 2 sticky ends. These plasmids consists of Agrobacterium tumefaciens ‘s DNA (Roller, 1998).
Then, the sticky ends are then mixed with plasmid DNA. Next, DNA ligase enzymes are necessary to rejoin the plasmids. However, these ligase enzymes can only be activated with specific conditions such as temperature. The rejoined plasmids are then reintroduced into the bacterium and the bacterium is cultured under standard microbial methods (Mohan, 2005).
The bacterium is then introduced into the plant, where the plant will grow a gall in reaction to the bacterium. A certain number of cells from the gall might contain the required insecticidal gene (Roller, 1998). These sections of the gall may be encouraged to grow by special plant tissue culture techniques, bulked up in the lab before conditions in the medium are changed to encourage growth of roots and shoots (Mohan, 2005). The resulting small plants are then transferred into the field, along with other crops.
author: monica
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