Lecture Handout 9 - BIOT 101 Lecture Handout 9 Recombinant...

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BIOT 101 Lecture Handout 9 Recombinant DNA Technologies Vocabulary autonomously replicating sequence (ARS) Auxotrophic mutants Bacterial artificial chromosomes (BACs) beta-lactamase bacteriophage vectors cDNA cDNA libraries centromere clonal population clones cloning technologies cloning vector copy number cos sites cosmid DNA construct electroporation foreign DNA genetic engineering genomic libraries insert DNA transgene Microinjection Microprojectiles multiple cloning site (MCS) mutagenesis origin of replication (ORI) phage lambda 1 plaques plasmid polylinker region protoplast recombinant DNA technologies reported gene selectable marker shotgun cloining Shuttle vectors Silicon-carbide fibers telomeres transformation transformed cells transgenics Yeast artificial chromosome (YACs) Objectives Understand the reasons for genetically engineering organisms. Be able to explain the steps that are undertaken in a genetic engineering project. Be familiar with the various cloning vectors that exist. Understand the limitations of these vectors. Understand the methods for introduction of DNA into cells. Understand how transformed cells are differentiated from nontransformed cells and why such differentiation is necessary. Understand what reported genes are and why these systems are necessary. 1
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BIOT 101 Lecture Handout 9 Recombinant DNA Technologies I. The goal of genetic engineering is to alter the DNA of an organism. This alteration may involve altering existing DNA ( mutagenesis ) or the introduction of DNA from other organisms ( transgenics ). The methodologies that allow for introduction of foreign DNA into an organism are referred to as recombinant DNA technologies or simply cloning technologies . A. Introduction of foreign DNA into a new host cells can be done for several reasons. 1. Researchers may need large amounts of DNA for such purposes as DNA sequencing or use as hybridization probes. In such a case, the DNA of interest will be introduced into cells in such a way that the cell will make multiple copies of the DNA. The genetically modified cells will be allowed to grow creating a large population of cells carrying the DNA of interest. The researcher then isolates the DNA from the recombinant cells. 2. Researchers may need large amounts of the protein for research or for commercial purposes (therapeutic proteins, industrial enzymes, etc.). In this case, the foreign DNA carries a gene for the desired protein and is introduced into cells in a manner that results in production of the protein by the recombinant cell. The recombinant cells are allowed to grow creating a large population of cells that are producing the protein of interest. The protein can then be purified from the culture or culture medium (fluid in which the cells were growing). 3.
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This note was uploaded on 12/07/2009 for the course BIOT 101 taught by Professor Don during the Fall '09 term at Ivy Tech Community College.

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Lecture Handout 9 - BIOT 101 Lecture Handout 9 Recombinant...

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