Presentation on theme: "Ch. 13.1: BIOTECHNOLOGY Objectives: 1.Explain how the use of bacteria has contributed to the development of DNA technology. 2.List some recent research."— Presentation transcript:
Ch. 13.1: BIOTECHNOLOGY Objectives: 1.Explain how the use of bacteria has contributed to the development of DNA technology. 2.List some recent research trends in recombinant DNA technology. Vocab: Biotechnology Recombinant DNA technology
Biotechnology = Use of organisms to perform practical tasks. Examples: Use bacteria to make proteins/drug therapies Engineer plants with natural resistance to insects, drought, fungi, etc. Develop vaccines.
Recombinant DNA = DNA from 2 different sources combined.
13.2: Bioengineering Vocab: PlasmidRestriction enzyme Genomic libraryNucleic acid probe Objectives: 1. Explain the role of plasmids in engineering bacteria. 2.Explain how biologist “cut and paste” DNA. 3.Describe the procedure used in cloning a specific gene. 4.Identify the usefulness of recombinant microorganisms.
Bacteria: Work horses of Biotech. Used to mass produce useful genes + proteins. Simple organisms 1 chromosome. Plasmids Reproduce rapidly Easy to manage in a lab.
Bacteria Plasmids Small, circular DNA Separate from chromosome. Contains a few genes. Make copies of itself Can be shared/ transferred b/w bacteria. Exchange of plasmids is how bacteria build genetic variation. Genes for antibiotic resistance are shared this way :(
Making Recombinant DNA 1.Remove desired gene from donor cell. 2.Remove bacteria plasmid. 3.Insert gene into plasmid. 4.Return plasmid to bacteria. 5.Gene is transcribed and translated into protein product. Recombinant DNA
Restriction Enzymes = Enzymes that CUT foreign DNA sequences. Each enzyme is cuts a specific sequence (CCCGGG or GAGCT) Cuts sugar-phosphate backbone of DNA Make staggered cuts. Leave “sticky ends” on cuts. Evolved in bacteria to protect bacteria fr. invading viruses.
Restriction Enzymes: Cut DNA into fragments Sticky ends = made by staggered cuts; unpaired bases; useful b/c they h-bond w/ complimentary bases in other fragments. Helps to “sew” fragments together & make recomb. DNA. BLUNT ends =no staggered cuts; not as useful
Ligase: Glues DNA fragments together
Genetic Cloning = Copies of recomb. DNA (and resulting proteins) are made by reproducing organisms
Genomic Library = Complete collection of cloned DNA fragments from an organisms. When you use restriction enzymes they cut up the donor DNA into MANY fragments. Each fragment is incorporataed into a plasmid. You need to figure out which bacteria has desired recomb. Plasmid!
Identifying Desired Recomb. DNA 1.Use nucleic acid probe Radioactively labeled complimentary sequence (TAGGCT will find and bind to ATCCGA when strands are separated). 2. Insert desired DNA into plasmid sequence for antibiotic resistance. Recomb. plasmids will lose resistance to antibiotics and will NOT survive when exposed to antibiotic.
Task: Diagram Steps in Creating recombinant DNA clones Use and illustrate the following key terms… Host DNA Bacteria Plasmid Vector Restriction Enzyme Sticky Ends Ligase Bacteria reproduces/plasmid replication Recombinant DNA Clones
Lab: Recombinant Paper Plasmids Goal: Insert human gene for insulin production into bacteria plasmid. 1.Construct bacteria plasmid. Color code sequences for antibiotic resistance. 2.Label 3’ and 5’ ends of restriction enzymes and plasmid DNA. 3.Identify restriction enzymes that will cut plasmid in sequence for antibiotic resistance. Mark these locations and label the enzyme used.
Lab: Recombinant Paper Plasmids Goal: Insert human gene for insulin production into bacteria plasmid. Label 3’ and 5’ of human DNA Determine the enzyme that will allow for removal of human insulin gene AND match up with the sticky ends on the plasmid. Create the recombinant plasmid.