gene encoding the enzyme lactate dehydrogenase [ldhA] from the bactaria was isolated and cloned. The expressed ldhA gene was inserted into pET28b plasmid vector. The resulting recombinant pET28b-LdhA expression vector was then transformed after introduction into E. coli. The ligation gave 15 colonies of recombinant DNA which later gave 6 recombinant plasmids as revealed by gel electrophoresis. 1. Introduction The NADH-dependent lactate dehydrogenase (EDH) is a key enzyme in the fermentative
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Consider the following double-stranded DNA sequence: 5’-CAG AAG AAA ATT AAC ATG TAA-3’ 3’-GTC TTC TTT TAA TTG TAC ATT-5’ If the bottom strand serves as the template‚ what is the mRNA sequence produced by transcription of this DNA sequence and Why? 5’-CAG AAG AAA AUU AAC AUG UAA-3’ mRNA sequence 3’-GTC TTC TTT TAA TTG TAC ATT-5’ DNA template strand We get the mRNA sequence due the transcription process‚ which gives us the RNA bases that are complementary to the DNA template
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1. Introduction Since the discovery of plasmid‚ various methods have been developed to isolate plasmid DNA. All the methods have one common and important target of isolating plasmid DNA of high quality and quantity in less time. These methods are not completely safe because of use of toxic chemicals compounds. The developed protocol for plasmid extraction is based on the alkaline lysis method of plasmid preparation (extraction at pH 8.0) with slight modifications. Cell lysis reagent sodium dodecyl
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Transformation__ competent cells required f. Conjugation ____ an F plasmid encodes attachment proteins g. Conjugation _____used to construct the E. coli minute map h. Conjugation_____ a pilus is produced i. Transduction____ lytic cycle entered in response to stress j. Transduction____ may be specialized or general k. Conjugation_____ requires cell to cell contact l. Transduction ____mediated by a virus
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This experiment is to determine the unknown DNA plasmid using restriction enzymes and conducting electrophoresis finally comparing the resulting fragments with the known restriction map. In this lab‚ it succeeds in showing the fragments. In this report we will discuss the‚ results‚ limitations and possible errors. Introduction In biology restriction enzymes are used in several ways to modify and manipulate DNA molecules. One common use is to compare pieces of DNA from one that is unknown‚ with fragments
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together are the complementary bases‚ or sticky ends. Question #3: What is a plasmid? How is a plasmid used in gene splicing? A plasmid is a circular segment of DNA with an origin of replication. It can be duplicated and serve as an adjunct to the main bacterial chromosome. Question #4: What types of vectors are used to carry DNA from one species into the DNA of another species? Give examples. Generally plasmids and viruses‚ although naked DNA is a possible source as well. Question #5:
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biological vectors like plasmids or viruses. Other newer gene transfer techniques are electro- and chemical poration‚ microinjection‚ and bioballistics. Recombinant DNA Recombinant DNA techniques use biological vectors like plasmids and viruses to carry foreign genes into cells. Plasmids are small circular pieces of genetic material found in bacteria that have the ability to cross species boundaries. The circles can be broken and new genetic material added to them. Plasmids augmented with new genetic
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molecules for the bacterium. The process begins when a plant is wounded‚ often near the base of the stem. This causes the release of compounds that attract Agrobacterium to the damaged cells. Transcription of genes on one of the bacterium’s plasmids begins‚ transferring a 20 kbase piece of single-stranded T-DNA into the plant nucleus. Here the T-DNA randomly integrates into the plant chromosome‚ translating 2 groups of substances:- 1. One group of chemicals causes an over-production of
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Vinciane Dufour Service de Biologie Moleculaire‚ Agence Française de Sécurité Sanitaire des Aliments (AFSSA)‚ Ploufragan‚ Brittany‚ France Introduction In 1990‚ Wolff and his colleagues were the first to report the successful expression of naked plasmid DNA in mouse muscle tissue [108]. A few years later it was reported that the injection of DNA encoding an antigenic protein of influenza virus conferred protective immunity in mice [97]. Many papers have described and reviewed the protective immunity
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KEY: ABE Lab 1: Prelab: Page A17‚ Questions 1 and 2: 1. What does the term genetic disease mean? What examples of generic diseases do you know about? A genetic disease is a disease caused by an absent of defective gene in the DNA. Students may know of sickle cell anemia‚ which is a disease resulting from defective red blood cells; treatment of sickle cell anemia focuses primarily on relieving pain and controlling infections and other complications that may occur. 2. Adding human DNA to bacteria
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