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Plasmids By: Brye, Hannah, and Jen What are plasmids? ● ● ● ● Genetic structure in a cell that can independently replicate chromosomes It is typically a small circular DNA strand in the cytoplasm of a bacterium Used mainly in the manipulation of genes Plasmids can be found in all three major domains, Archaea, Bacteria and Eukarya. Importance of Plasmids ● ● ● ● Easy to work with Self-replicating Stable Functional in many species and can be useful for a diverse set of applications Plasmids play a very important role in genetic engineering, particularly in gene cloning. When genes are cloned, the process usually takes place within bacteria. In order to get the gene that is to be cloned into the bacteria, a vector is necessary. A plasmid is what is used as the vector, as it can pass easily from one cell to another. Biotechnology ● ● ● ● ● ● ● Plasmids have been key to the development of molecular biotechnology. They act as delivery vehicles, or vectors, to introduce foreign DNA into bacteria. Using plasmids for DNA delivery began in the 1970s when DNA from other organisms was first ‘cut and pasted’ into specific sites within the plasmid DNA. The modified plasmids were then reintroduced into bacteria. Decades after their first use, plasmids are still crucial laboratory tools in biotechnology Scientists can force bacteria to keep them. Virtually all plasmids that are used to deliver DNA contain genes for antibiotic resistance. Once bacteria have been treated with a plasmid, scientists grow them in the presence of antibiotic. Only those cells that contain the plasmid will survive, grow and reproduce. The others will be killed by the antibiotic. They are copied independently. Plasmids can be copied numerous times, regardless of whether the bacterial host is replicating its own DNA, and every time a plasmid vector is replicated, so is the introduced DNA that it contains. They are circular. DNA that is circular is well suited to incorporate extra DNA sequences. That’s because it can be cut open without falling apart, then snap back together once new DNA has been incorporated. Major steps involved Steps in cloning a gene: 1. 2. 3. 4. 5. Plasmid DNA is isolated DNA containing the gene of interest is isolated Plasmid DNA is treated with a restriction enzyme that cuts in one place, opening the circle DNA with the target gene is treated with the same enzyme and many fragments are produced Plasmid and target DNA are mixed and associated with each other Applications in the medical field and Usage Strategies utilizing plasmid DNA have been used for non-treatable neurological disorders, such as Parkinson disease (PD), Alzheimer disease (AD), and multiple sclerosis (MS). Because of their ability to move genes from cell to cell, plasmids have become resourceful tools for both research and biotechnology. In the laboratory, researchers use plasmids to carry marker genes, allowing them to trace the plasmid’s inheritance across host cells. Transferred or cloned genes are used to produce a variety of important medical, agricultural, or environmental products that can be economically used by humans. Unknowns ● ● ● ● ● ➔ ➔ Naturally occurring stable elements which are found in bacteria, fungi and even bacteria of some plants Composed of dna or rna, single or double stranded, linear or circular Exist and replicate independently of the chromosome in which they are found Not required by their host cell for survival Subgrouped into 5 main types based on their function R plasmids: carry genes encoding resistance to antibiotics Col plasmids: confer on their host for the ability to produce antibacterial polypeptides called bacteriocins that are often lethal to closely related or other bacteria Unknown continued ➔ ➔ ➢ ➔ ➔ The col proteins of E. coli are encoded by plasmids such as ColE1. F plasmids contain the F or fertility system required for conjugation (the transfer of genetic information between two cells). These are also known as episomes because, under some circumstances, they can integrate into the host chromosome and thereby promote the transfer of chromosomal DNA between bacterial cells. Degradative or catabolic plasmids allow a host bacterium to metabolize normally undegradable or difficult compounds such as various pesticides. Finally, virulence plasmids confer pathogenicity on a host organism by the production of toxins or other virulence factors. Cool Facts ● ● ● ● Plasmid size varies from 1 to over 1,000 kilobase pairs (kbp) The number of identical plasmids within a single cell can range from one to thousands Extremely effective cloning vector Plasmids may carry genes that provide resistance to naturally occurring antibiotics in a competitive environmental niche Ethical/ Moral issues ● ● ● ● ● Depends on what the plasmid contains, some have origins of replication which mean they can replicate independently of the cell nucleus, they can contain genes that confer a selective advantage There's always controversy over the subject of cloning Some people believe that there should only be one of each individual Many scientists believe that it is “playing God” Some believe it is unnatural Video https://www.youtube.com/watch?v=GNMJBMtKKWU Referencing https://www.addgene.org/mol-bio-reference/plasmid-background/ https://www.ncbi.nlm.nih.gov/pubmed/22023479 http://www.encyclopedia.com/science-and-technology/biology-and-genetics/genetics-and-genetic-en gineering/plasmid http://www.wisegeek.com/what-are-plasmids.htm http://www.encyclopedia.com/science-and-technology/biology-and-genetics/genetics-and-genetic-en gineering/plasmid http://biotechlearn.org.nz/themes/bacteria_in_biotech/bacterial_dna_the_role_of_plasmids