Introduction to Biotechnology Manipulating and Analyzing DNA

Uses of Biotechnology Analyzing human DNA DNA fingerprinting Genetic diseases DNA fingerprinting Forensics The Human Genome Project Identifying all genes in the human genome Gene therapy Curing disease

Genetic Engineering Definition: introducing a set of genetic information (genes) directly into an organisms DNA How is this done? With use of restriction enzymes Plasmid DNA

Restriction Enzymes Proteins originally made to protect against viruses Restriction Enzymes chut DNA at specific sequences called recognition sites Recognition sites are sequences of DNA that enzymes recognize

The “Ends” Two types of cuts result from restriction enzymes: “Sticky” ends leave exposed bases ready to hydrogen bond. Eco RI below is an example. “Blunt” ends leave NO bases exposed Eco RV below is an example DNA ligase joins “the ends” of cut DNA together

DNA Recombination DNA fragments cannot function all by themselves They must become part of the genetic material of LIVING cells before the genes they contain can be activated DNA fragments + DNA from a living cell = Recombinant DNA DNA fragments are often inserted in plasmids (found in bacteria) that have been cut with the same sticky ends

Plasmids Circular double-stranded DNA (bacterial) Used in biotechnology because they are circular, small, and replicate on their own

Plasmids + DNA of interest Restriction enzyme EcoRI used to isolate gene of interest Same restriction enzyme EcoRI used to cut plasmid DNA Sticky ends match up. Gene of interest is inserted into plasmid=recombinant DNA

Genetic Engineering Example Insulin is made for diabetics a) Cut open plasmid and DNA with gene encoding for insulin with restriction enzymes b) Mix cut plasmid with cut DNA (they have the same sticky ends) c) Seal with ligase d) Insert recombinant DNA (plasmid) into bacteria e) Recombinant DNA replicates and bacteria divides f) DNA is transcribed and translated = insulin

Engineering New Organisms Transgenic Organisms: Organisms that contain “foreign” DNA are termed transgenic May be bacteria, plants, or animals

Examples of GFP gene

This All Involves One Thing… DNA Sequencing: reading or identifying the sequence of bases along the length of a DNA molecule AND understanding what that sequence codes for…