Transgenesis and Gene Cloning

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Transgenesis and Cloning Transgenesis is the process of inserting a gene from one source into a living organism that would not normally contain the inserted gene. The gene can come from the same species (called Cisgenesis) or from a different species entirely. To facilitate the transfer of genes from one organism to another, often a Transgenic Organism with Recombinant DNA is created: -The first step in creating an organism capable of carrying out the transformation process is to isolate the required gene. This is done so using Restriction Enzymes, which target a specific gene sequence. The gene is often cut with staggered ends, called “Sticky Ends” which only allow specific and complementary gene sequences bond by base pairing. Due to these “Sticky Ends”, the scientists can exercise a degree of control over where the genes bond. -The next step in the process is to amplify the gene of interest using Polymer Chain Reaction or PCR. PCR serves to replicate specific gene sequences, creating many copies for the scientists to work with. -After the gene is amplified a suitable vector is selected for use. A vector is a self-replicating DNA molecule used to transmit a gene from one organism to another. All vectors must contain the following characteristics: 1. Able to replicate host organism. 2. One or more sites that a restriction enzyme can cut. 3. Have some kind of genetic markers that allows them to be easily identified. Examples of vectors used are yeast organisms, viruses and bacterial plasmids. Each has their own advantages and disadvantages. In this case bacterial plasmids are used. -The plasmid is then extracted and cut using the same restriction enzyme used to cut the gene of interest. This is to create ends that are complementary to each other. Often the scientists cut the plasmid at specific places, often disrupting a specific characteristic. For

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