Chapter 20 outline - Chapter 20: h h h DNA technology makes...

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Unformatted text preview: Chapter 20: h h h DNA technology makes it possible to make recombinant DNA recombinant Recombinant DNA is DNA in which genes Recombinant from two sources are combined into the same molecule same Allows for genetic engineering, or Allows manipulation of genes. manipulation DNA technology has spawned the DNA field of bio technology to make useful products. useful Bacterium Gene inserted into plasmid Cell containing gene of interest h h Gene cloning makes it easier to study specific parts. specific Genes can be cloned by inserting them into Genes bacterial plasmids bacterial The copied genes can either be studied The individually or researchers can massindividually synthesize proteins for study Recombinant DNA (plasmid) Gene of interest Plasmid put into bacterial cell DNA of chromosome Recombinant bacterium Host cell grown in culture to form a clone of cells containing the “cloned” gene of interest Gene of interest Protein expressed by gene of interest Copies of gene Protein harvested Basic research on gene Basic research and various applications Basic research on protein Bacterial chromosome Plasmid Gene for pest resistance inserted into plants Gene used to alter bacteria for cleaning up toxic waste Protein dissolves blood clots in heart attack therapy Human growth hormone treats stunted growth h h h t Restriction enzymes make genetic engineering possible. engineering Protects DNA from intruder DNA Have recognition sequences to be cut out of Have DNA DNA Results are sticky ends Results sticky New DNA can be fused to these sticky ends with DNA ligase. h h h Gene is isolated and put into cloning vector. Gene Insert cloning vector into cells Clone cells by putting them on a nutrient Clone rich medium containing ampicillin and X-gal sugar. sugar. Identify cells containing genes of interest h h h Scientists employ an expression vector – expression a cloning vector that contains prokaryotic promoters which can express the foreign gene inserted. gene Complimentary DNA (cDNA) – DNA Complimentary containing sequence for gene, but no introns. introns. Yeast Artificial Chromosomes (YAC) can be a eukaryotic vector for can gene cloning. gene Electroporation h h Alternate way to clone a gene if source DNA is in poor condition. is Technique to amplify any piece of DNA Technique without using cells. without The DNA is put in a test tube with a special The kind of DNA polymerase. The procedure consists of heating DNA to separate it and let it form copies. form h h Compares DNA sequences because it separates macromolecules on basis of size. separates Electricity strains the macromolecules Electricity through the gel. The molecule’s rate of movement mostly depends on its length. movement With the current off, the DNA is arranged in With a neat column from largest to neat smallest. h h Method to compare DNA between 3 individuals. individuals. Valuable because it shows differences in Valuable noncoding DNA as well as coding DNA. noncoding How it’s done Complementary strands that have been Complementary radioactively labeled attach to genes radioactively of interest. Photogenic film shows only labeled genes to compare only certain genes. h h h h Human Genome Project, possible by DNA technology. technology. Effort to map entire human genome by Effort ultimately determining the complete nucleotide sequence for each chromosome in the body. in To map, they create a genetic linkage map. They physically map as well by observing They gene distances. gene They utilize Bacterial artificial chromosomes They as well as Yeast artificial chromosomes. as Sanger method – instead of Human Genome project. Genome Synthesized strands complementary to the DNA Synthesized being sequenced and pieced them together with a powerful computer program. powerful h h Scientists scan DNA for signs of proteincoding genes. I.E. Absence of repetitive coding DNA. DNA. Humans are estimated to have about 30,000~40,000 non-repetitive genes. h h We study genes not only to ascertain their functions, but also to learn how genes work together. together. DNA Microarray Assays – tiny amounts of a DNA large number of single stranded DNA fragments representing different genes are fixed in a grid. fixed Tested for hybridization with different enzymes and see if they activate. h in vitro mutagenesis – force a gene to mutate so that they can look for changes in the phenotype. the RNA interference (RNAi) – Uses a synthetic RNA double banded RNA matching the sequence of a particular gene to trigger breakdown of the gene’s mRNA. the Theory to be self defense against viruses that try to move retrotransposons. h After genomics, Scientists will pursue proteomics for proteins. h h Bioinformatics – Computer science and mathematics to aid in dealing with the large amount of data. amount Single nucleotide polymorphisms – Seem to Single be how we are differentiated in the overall genome. genome. We are probably 99.9% identical DNA – wise. Diagnosis of diseases Diagnosis Track down pathogens such as HIV Can identify genetic disorders before the Can problems onset. problems Detect abnormal genes present in DNA. h Gene Therapy Alter afflicted genes Must multiply throughout Must patient’s life, such as stem or bone patient’s marrow cells. Gene Therapy Gene Safety concerns limit use to deadly defects Need to make sure they don’t harm another Need cell’s function somehow. cell’s Moral issues Interference of Evolution h Pharmaceutical Products Expression vectors make large amount of proteins. Block cell receptors (for HIV) Stimulate immunity by producing protein found on pathogen. Forensic use Forensic Southern Blotting useful to compare DNA of Southern suspects. suspects. DNA fingerprint – specific pattern of bands. Compare simple tandem repeats Compare (microsatellites) (microsatellites) Very reliable h Agricultural Use Transgenic organisms – contain genes from other species. Genetic engineering; human blood protein. h Provide crops with desirable traits by genetically modifying them. genetically Ti plasmid – vector used for plant cells from Ti the soil bacterium Agrobacterium tumefaciens. tumefaciens A drawback is that sometimes the bacterium drawback infects the DNA… infects h h Quickly reducing plant breeding programs. Trying to increase nitrogen fixation to stop fertilizer use. h h h h h Worried about creating hazardous pathogens for warfare. pathogens Dangerous Experiments banned Worries about genetic modified organisms Enhanced crops may be harmful to humans Rise of “Superweeds” Ethical questions with human genome. ...
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Chapter 20 outline - Chapter 20: h h h DNA technology makes...

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