Hybridization of the Plasmid and Foreign DNA - Essay Example

The latter permits replication of the plasmid in host cells. At the same time, the drug-resistance gene is essential in allowing host cell growth by destroying antibiotics, especially carbenicillin. The use of restriction enzymes in obtaining cloning sites through the cleaving of the vector acts as the initial step in inserting foreign DNA (Seviour and Nielsen, 2010, p. 365). Sticky ends, especially single-stranded ends, can result from cleavage of palindromic sequence GAATTC by EcoRI (Russell et al., 2013, p. 393). The resultant single-stranded ends have hybridization ability with similar EcoRI pieces of DNA. Using the principle, scientists take sequences of foreign DNA for cloning and mix them with cleaved vectors after digestion with EcoRI (Brown, 2013).

After the hybridization of the plasmid and foreign DNA through the sticky ends, the next step involves sealing with phosphodiester linkages to form a recombinant plasmid. DNA ligase enzyme remains imperative in the sealing process. Consequently, the replication above the origin, resistance gene, and DNA fragment stay on the newly created recombinant plasmids that collectively form a circular library. The inherent recombinant plasmids have each of them possessing a unique foreign DNA fragment. The subsequent stage involves the addition of E. Coli bacteria that act as host cells to the recombinant plasmids. At this stage, the study has made the cells permeable to DNA through treatment with CaCL2. Some cells resist taking recombinant plasmids, while others do through a process known as transformation.

The researcher pours the E Coli cells into an antibiotic carbenicillin plate nutrient agar. It is important to note that only cells resistant to carbenicillin in antibiotics would grow in the agar as opposed to the rest. As mentioned above, the growth and multiplication of the cells remain possible at 37 degrees Celsius. Lack of movement of the cells within nutrient agar causes the formation of cell colonies in the plate. The construction of settlements occurs separately in the media. The cells that have died did not get transformed, lack plasmid, and will not resist the antibiotic carbenicillin.

Consequently, the non-transformed cells would die because of lacking the resistance gene. However, the transformed cells have a plasmid that can resist the antibiotic and will survive. During cell division, plasmids get distributed to daughter cells despite their independent replication mechanism. During replication and subsequent multiplication of the host cells, there would exist amplification of the recombinant plasmids. The results are a clone or multiple colony daughter cells. The resultant host cells have similar recombinant plasmid copies and foreign DNA fragments mainly because of derivation from a single cell.