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TAMU BIOL 111 - Chapter 20 PPT Notes

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Chapter 20 PPT Notes1. Biotechnology: manipulation of organisms or their components to make useful products2. Applications of DNA technology affect everything from agriculture, to criminal law, to medical research3. Nucleic acid hybridization: the base pairing of one strand of nucleic acid to the complementary sequence on another strand4. Genetic engineering: direct manipulation of genes for practical purposes5. DNA sequencing: exploit the principle of complementary base pairing to determine a gene’s complete nucleotide sequencea. “next-generation sequencing” techniques use single template strand that is immobilized and amplified to produce an enormous number of identical fragments6. DNA cloning: scientists prepare well-defined DNA segments in multiple identical copies by this process7. Plasmids: small circular DNA molecules that replicate separately from the bacterial chromosome 8. Recombinant DNA: molecule with DNA from two different sources9. Gene cloning: production of multiple copies of a single gene is a type of DNA cloninga. Useful for amplifying genes to produce a protein product for research, medical, or other purposes10. Cloning vector: plasmid used to clone a foreign genea. Bacterial plasmids widely used as cloning vectors because they’re readily obtained, easily manipulated, easily introduced into bacterial cells, and once in the bacteria they multiply rapidly11. Restriction enzymes: cut DNA molecules at specific DNA sequences called restriction sites12. Restriction fragments: the cuts a restriction enzyme makes13. “Sticky ends”: fragments produced by the most useful restriction enzymes that cut DNA in a staggered way14. DNA ligase: enzyme that seals the bonds between restriction fragments15. Gel electrophoresis: technique uses a gel made of a polymer to separate a mixture of nucleic acids or proteins based on size, charge, or other physical propertiesa. Negatively charged DNA move toward the positive electrode, smaller fragments move faster and further than larger ones16. Polymerase Chain Reaction (PCR): produces many copies of a specific target segment of DNAa. Taq polymerase used because it is a heat-stable DNA polymeraseb. 3 steps (heating, cooling, and replication)17. Expression vector: cloning vector that contains a highly active bacterial promoter18. Eukaryotic DNA cloning and expression systems: can avoid eukaryote-bacterial incompatibility issues by using eukaryotic cells, such as yeasts, as hosts for cloning and expressing genes19. Electroporation: applying a brief electrical pulse to create temporary holes in plasma membranes (method of introducing recombinant DNA into euk cells)a. Scientists can inject DNA into cells using microscopically thin needles, once in thecell, the DNA is incorporated into the cell’s DNA by natural genetic recombination20. mRNA can be detected by nucleic acid hybridization with complementary moleculesa. molecules of either DNA or RNA are nucleic acid probes21. In situ hybridization: uses fluorescent dyes attached to probes to identify the location ofspecific mRNAs in place in the intact organism22. Reverse transcriptase-polymerase chain reaction (RT-PCR): useful for comparing amounts of specific mRNAs in several samples at the same timeda. Reverse transcriptase is added to mRNA to make complementary DNA (cDNA): serves as a template for PCR amplification of the gene of interestb. Products ran on a gel and mRNA is identified23. DNA microarray assays: compare patterns of gene expression in different tissues, at different times, or under different conditionsa. By uncovering gene interactions and clues to gene function DNA microarray assays may contribute to understanding of disease and suggest new diagnostic targets24. In vitro mutagenesis: mutations are introduced into a cloned gene, altering or destroying its functiona. When mutated gene is returned to cell, the normal gene’s function might be determined by examining the mutant’s phenotype25. RNA Interference (RNAi): the silencing of gene expression26. SNPs (single nucleotide polymorphisms): single nucleotide variants, are among the most useful genetic markersa. SNP variants found frequently associated with a certain inherited disorder let researchers know its’ likely location for disease-causing geneb. SNPs are rarely directly involved in the disease; they’re mostly in noncoding regions of the genome27. Stem cell: relatively unspecialized cell that can reproduce itself indefinitely, or under certain conditions can differentiate into one or more types of specialized cells28. Totipotent cell: cell that can generate a complete new organism29. Nuclear transplantation: the nucleus of an unfertilized egg cell or zygote is replaced withthe nucleus of a differentiated cell30. Pluripotent: embryonic stem (ES) cells are capable of differentiating into many different cell typesa. Ultimate aim of research with stem cells is to supply cells for the repair of damaged or diseased organs31. Induced pluripotent stem cells (iPS): cells used as models for study of certain dieseases and potentially as replacement cells for patients; can perform most of the functions ES cells can perform32. Transgenic: animals made by introducing genes from one species into the genome of another animal33. STRs are variations in the number of repeats of specific DNA sequencesa. PCR and gel electrophoresis are used to amplify and then identify STRs of different


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