myguide exam 4

myguide exam 4 - Study Guide Exam#4 Chapter 13...

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Study Guide Exam #4 Chapter 13 Biotechnology Vocab – Biotechnology – the application of organisms, biological cells, cell components, and biological processes to practical operations and procedures. Recombinant DNA – a form of DNA from one species that has been modified by incorporation of a DNA segment from another species; hybrid DNA Plasmid – self-replicating, extrachromosomal DNA molecules found in bacteria; contain genetic information for the translation of proteins that confer a specialized characteristic; used as cloning vectors Cohesive ends Transformation – the introduction of hybrid DNA into a host organism, in which it can be replicated Genomic library – a collection of bacterial or yeast cells that have been transformed using recombinant vehicles with DNA inserts from a single species. Polymerase chain reaction – a laboratory method used to synthesize amplified quantities of specific nucleotide sequences of DNA from small amounts of DNA using heatstable DNA polymerase and cycles of denaturation and replication. RFLPs – restriction fragment length polymorphisms – a method used for the analysis of DNA fingerprinting data; uses restriction endonucleases to cleave the DNA and electrophoresis to compare fragments from different individuals. Gene therapy – medical procedures used to correct a genetic defect by inserting the normal gene into the cells of an organism Blotting – transfer of molecules (especially recombinant DNA) from gels to paper for genetic analysis. Vector – an agent that serves as the carrier for foreign DNA in recombinant DNA technology; also called a vehicle Transgenic organisms – an organism whose genetic material has been altered using genetic engineering techniques known as recombinant DNA technology. DNA ligase – class of enzymes that catalyzes bond-forming reactions using energy from ATP Study guide Steps in basic molecular cloning using a plasmid and E. coli as host including transformation and selection of transformed cells based on antibiotic resistance 1. select and isolate a DNA molecule to serve as the carrier (vector) for foreign DNA 2. cleave deoxyribonucleotide strands of the vector (DNA carrier) with a restriction endonuclease 3. Prepare and insert the foreign DNa fragment into the vector. This produces a recombinant of hybrid DNA 4. Introduce the hybrid DNA into a host organism (bacterial cell usually) where it can be replicated. This is called transformation 5. Develop a method for identifying and screening the host cells that have accepted and are replicating hybrid DNA 6. Purify the protein product Additional criteria for an expression vector – used to produce a desired protein 1. plasmid should replicate in a relaxed fashion so that many copies are produced
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2. the plastmic should be small; then it is easier to separate from the larger chromosomal DNA, easier to handle without physical damage, and probably contains only a few sites for attack by restriction endonucleases 3. The plasmid should contain identifiable markers for screening progeny for the presence of the
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myguide exam 4 - Study Guide Exam#4 Chapter 13...

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