AP Bio Prelab 9


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THE PROCESS BEING USED IN THE EXPERIMENT Introduction: Plasmids- small circular pieces of DNA in prokaryotes Exist outside the Chromosome Some exist only as single copies in a bacterial cell and replicate only when the chromosome replicates Some replicate autonomously and occur in copies of 10-200 R Plasmids- special plasmids carrying genes for resistance to antibiotics Escherichia coli is an ideal organism for genetic manipulation and has been used in recombinant DNA research Commonly inhabits the human colon Can easily grow in a suspension culture in a nutrient medium (Luria Broth) or a petri dish of Luria Broth mixed with LB Agar or Nutrient Agar An E. coli chromosome contains 5 million DNA base pairs (1/600 th of a human’s DNA) as well as numerous Plasmids Genes can naturally be transferred through Conjugation, Transduction, or Transformation, and can give traits to an organism that it previously lacked Conjugation- a mating process in which genetic material is transferred from one bacterium to another (one-way) Transduction- a Bacteriophage (virus) carries genes between bacterium Transformation- genetic information is directly taken into the cell from the environment o Natural occurrences are extremely rare Only occur when bacteria are Competent- a period at the end of logarithmic growth when bacteria can accept external DNA Competence can be induced using chemical growth reactions Plasmids can transfer genes that occur naturally within them, and can introduce foreign DNA from other bacteria, plasmids, or eukaryotes Restriction Endonucleases can cut and insert foreign DNA pieces into Plasmid vectors Plasmid DNA usually contains genes for MULTIPLE traits Genetic Transformation involves inserting new DNA into E. coli cells Genetic Engineering- the process of inserting genes coding for new traits into Plasmids *Pertaining to the Experiment* The pGLO Plasmid carries the GFP gene that codes for green fluorescent proteins, and a gene that codes for a protein that gives resistance to the Ampicillin Antibiotic o GPF originates in the Aequorea victoria, a bioluminescent jellyfish o The beta-lactamose gene (provides resistance to ampicillin) is produced and secreted by bacteria containing the plasmid
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This note was uploaded on 12/28/2010 for the course BIO 103 taught by Professor Staff during the Fall '10 term at Ill. Chicago.

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