Chapter 14 Genetic Engineering Choose the best answer for each question. 1. Using this key‚ put the phrases in the correct order to form a plasmid carrying the recombinant DNA. Key: 1) use restriction enzymes 2) Use DNA ligase 3) Remove plasmid from parent bacterium 4) Introduce plasmid into new host bacterium. A. 1‚ 2‚ 3‚ 4 C. 3‚ 1‚ 2‚ 4 B. 4‚ 3‚ 2‚1 D. 2‚ 3‚ 1‚ 4 2. Which is not a clone? A. a colony of identical bacterial cells B. identical
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other to extract the DNA and RNA. After extracting the DNA‚ we it is important to use PCR amplification in order to amplify the DNA template to produce a specific DNA fragment. Another important step in DNA cloning is plasmid isolation. Plasmid isolation allows us to extract a plasmid from a bacterial cell (E.coli). In our experiments‚ we had to amplify either the 18S rRNA or the actin gene found in D. Melanogaster. Actin is a major contractile protein found in all eukaryotic cells‚ accounting
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METHODS In this experiment‚ plasmid lux and a control plasmid (pUC18) will be introduced into E. coli by transformation. There are four basic steps to the procedure; Preparation of competent cells (These steps should be performed by the instructor.) 1. Place a vial of CaCl2 solution and the tube of E. coli in the ice bath. 2. Using a sterile pipet‚ transfer 590µL CaCl2 solution to the tube containing 50µL of the bacteria. 3. Tap the vial with the tip of your index finger to mix the solution. 4. Incubate
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My research shows the rise in antibiotic resistant pathogens through horizontal gene transfer. Located in the bacteria are plasmids. They are independent‚ self-duplicating‚ and allow bacteria to perform new functions/generate new products. Basically plasmids help their hosts to stop the action of antibiotics and become resistant. “Gene transfer must be integral and critical to the overall survival of bacteria‚ providing a way for them to adapt to difficult conditions” (Levy 2002‚ 83). Horizontal
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the bacteria after you’ve changed it. • We can mess around with their DNA and kill a lot of them during our experiments and nobody gets mad. ;-) Now for a little vocabulary… Small‚ circular pieces of “extra” DNA found in bacteria. Plasmids Plasmids often carry antibiotic resistance. Restriction Enzymes: Molecular Scissors A restriction enzyme (RE) is a specialized protein that cuts DNA in a very specific place. • Different REs cut at different places along the nucleotide sequence
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of different origins produced in this way is called recombinant DNA‚ because it is DNA that has been recombined from different sources The combined techniques of using restriction enzymes and ligation are the basic tools of genetic engineering Plasmid is a circular piece of DNA from bacteria Two pieces of DNA are cut using restriction enzymes.
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want to study from the DNA strand . Then you would attach a target gene to a small‚ circular piece of DNA. Together‚ this is called a plasmid‚ which serves as the vehicle for transporting the gene. The nest step would be to put the plasmid into an E. coli cell or another type of bacteria. As each E. coli cell divides‚ each new cell contains a copy of the plasmid containing the gene. After that process it would grow a lot of E. coli cells Once your E. coli population has reached your desired number
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BACTERIA Period: 4 Characteristics: 3 major shapes Cocci Basilli Spirilla 3 major components Mesosomes flagella Plasmids Growing Up: Bacteria can obtain energy through phototrophs(sunlight)‚ lithotrophs(inorganic compounds)‚ and organotrophs(organic compounds) Marriage/Reproduction Binary Fission: The process by which all bacteria reproduce. It results in the separation of a single cell into two. Transformation: genetic alteration
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mut-goi from other components. A purified digested gene product is more advantageous for the DNA ligation‚ in which the gene products with sticky ends will be inserted to a plasmid vector. Also‚ to transform E.coli DH5α cells by introducing the plasmids DNA which contains the gene of interests into the E.coli strain(DH5α). The plasmid DNA can replicate inside the transformed E.coli DH5α cells‚ only successful transformed cells can produce the protein that is resistance to kanamycin‚ this allows for the
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we performed a genetic transformation through the process of gene transfer. Gene transfer involves the insertion of a gene into an organism. The gene to be inserted is usually contained in a plasmid‚ which is relatively small‚ circular non-chromosomal DNA molecule typically found in bacteria. Once the plasmid containing the gene is inserted into the organism‚ it is absorbed into the organism’s own genetic code. After this occurs‚ the newly introduced gene begins coding for proteins‚ giving the organism
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