DNA Gel Electrophoresis Research
Uploaded by spootyhead on Mar 05, 2007
DNA Gel Electrophoresis Research
The main objective of this lab was to identify unknown plasmids by observation of their genotype and phenotype. We observed the plasmids phenotype by using it to transform bacteria. When the plasmid is in a bacterial host the antibiotic resistance gene can be expressed and impart resistance to the host. We also analyzed the genotype of the unknown plasmid by performing a variety of molecular genetic techniques such as predigestion, electrophesis in argose gel, staining with ethidium bromide, and finally a photograph of the gel, in order to compare the plasmid to a standard, to in turn deterimine the plasmids actual size and the sizes of its digestion fragments. By performing these experiments we found that our unknown plasmid in the blue tube was resistant to pKAN. We also found that the unknown plasmid when compared to a standard closely matched the size of digestion fragments, of pKAN, therefore our unknown plasmid was pKAN.
Introduction
There is growing concern that the control of infectious diseases is threatened by the upward trend in the numbers of bacteria that are resistant to multiple antibiotics in the medical armamentarium. Resistance costs money and human lives. Resistant infections are associated with increased morbidity, prolonged hospital stays, greater direct and indirect costs, prolonged periods during which individuals are infectious, and greater opportunities for the spread of infection to other individuals (2). In many developing countries, the availability and use of antibiotics are poorly controlled, which results in a high rate of resistance, particularly to the older antibiotics (2).
The procedure used in our interpretation of Bacterial Transformation, is one that has a larger impact than our simple usage for finding unknowns when compared to knowns. This importance lies mainly in the medical field and more specifically in gene therapy. Using procedures, like this, Many human genes have been cloned in Escherichia coli or in yeast (4). This has made it possible - for the first time - to produce unlimited amounts of human proteins in vitro. Cultured cells (E. coli, yeast, mammalian cells) transformed with the human gene are being used to manufacture: insulin for diabetics, human growth hormone (GH), erythropoietin (EPO) for treating anemia, granulocyte-macrophage colony-stimulating factor (GM-CSF) for stimulating the bone marrow after a bone marrow transplant, adenosine deaminase (ADA) for treating some forms of...