<p>Duva’s Reading Guide for Chapter 13: Biotechnology</p><p>Concept 13.1: Recombinant DNA Can Be Made in the Laboratory</p><p>I. Recombinant DNA -</p><p>A. Restriction enzymes</p><p>A.1. Origin</p><p>A.2. Function</p><p>B. Recognition sequence/restriction site –</p><p>B.1. Blunt –</p><p>B.2. Sticky –</p><p>B.2.a. BamHI</p><p>B.2.b. HindIII</p><p>B.2.c. EcoRI</p><p>B.3. Palindrome</p><p>C. Modification of bacterial DNA</p><p>D. Restriction digest</p><p>II. Gel electrophoresis separates DNA fragments</p><p>A. Gel electrophoresis A.1. Purpose</p><p>A.2. Design</p><p>A.2.a. Gel</p><p>A.2.b. Buffer</p><p>A.2.c. Electric field</p><p>B. How does it work? C. Information obtained from the DNA sample</p><p>C.1. Number of fragments</p><p>C.2. Sizes of fragments</p><p>C.3. Relative abundance of a fragment</p><p>III. Recombinant DNA can be made from DNA fragments</p><p>A. DNA ligase</p><p>B. Cohen and Boyer</p><p>B.1. Bacterial plasmid</p><p>B.2. Results</p><p>Concept 13.2: DNA Can Genetically Transform Cells and Organisms I. Clone</p><p>II. Transformation</p><p>III. Transgenic</p><p>IV. Selectable marker</p><p>V. Model organisms for research</p><p>A. Bacteria</p><p>A.1. Pros</p><p>A.2. Cons</p><p>B. Yeasts</p><p>B.1. Pros</p><p>B.2. Cons</p><p>C. Plant cells</p><p>C.1. Pros</p><p>C.2. Cons</p><p>D. Cultured animal cells</p><p>D.1. Pros</p><p>D.2. Cons</p><p>VI. Methods for inserting DNA into host cells</p><p>A. Chemical treatment</p><p>B. Electroporation C. Use of viruses and bacteria as vectors</p><p>D. Inject DNA into nuclei of fertilized eggs</p><p>E. “gene guns”</p><p>VII. Replicon – </p><p>A. Can be inserted into host chromosome randomly</p><p>B. Vector</p><p>B.1. Plasmids</p><p>B.1.a. Small</p><p>B.1.b. Restriction enzyme recognition sites</p><p>B.1.c. Selectable marker (antibiotic resistance)</p><p>B.1.d. Replicate independently of host chromosome</p><p>B.2. Plasmid vectors for plants</p><p>B.2.a. Agrobacterium tumefaciens</p><p>B.2.b. Plasmid name</p><p>B.2.c. How does it work?</p><p>B.3. Viruses B.3.a. Bacteriophage λ</p><p>B.3.b. Pros</p><p>C. Reporter genes</p><p>Concept 13.3: Genes and Gene Expression Can Be Manipulated</p><p>I. Sources of DNA fragments</p><p>A. Genomic library</p><p>B. cDNA libraries</p><p>C. synthetic DNA and PCR</p><p>II. DNA mutations can be made in the lab</p><p>III. Genes can be inactivated by homologous recombination</p><p>A. Inactivate genes to understand their function</p><p>B. Stem cells</p><p>Concept 13.4: Biotechnology Has Wide Applications</p><p>I. Protein factories</p><p>A. Expression vectors</p><p>A.1. Prokaryotes</p><p>A.2. Eukaryotes</p><p>II. Medically useful proteins</p><p>A. Human insulin</p><p>A.1. Engineered plasmid A.2. Bacteria factories</p><p>A.3. Chemical treatment</p><p>B. Pharming</p><p>B.1. Protein produced in milk</p><p>B.2. HGH</p><p>III. Agriculture – transgenic plants</p><p>A. Produce insecticides</p><p>B. Improved nutrition</p><p>C. Adapt to environment</p><p>IV. Public concern</p><p>A. Unnatural</p><p>B. Unsafe</p><p>C. Dangerous to the environment</p>