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Biotechnology The use of biological processes, organisms, or systems to manufacture products intended to improve the quality of human life. Transgenics • Genetic engineering makes it possible to transfer DNA sequences from one organism to another • REMEMBER- DNA is universal • A gene taken from one species can be replicated in another species. Recombinant DNA • Recombinant DNA contains DNA from 2 or more different sources. • So how does DNA get from one species to another? • Vector: something used to transfer DNA from one organism to another – Microinjection – Gene Gun – Virus – Bacteria Plasmid Bacteria as vectors • Bacteria are used because they have plasmids • A plasmid is a circular piece of DNA that exists apart from the chromosome and replicates independently of it. • So how do I take a gene from one organism and put it into another? • We have to cut it using a Restriction Enzyme What are Restriction Enzymes? • Proteins • In nature: Used by bacteria to cut viral DNA • They “restrict” the growth of viruses • http://www.dnalc.org/resources/animatio ns/restriction.html Restriction Enzymes • Cut strands of DNA at specific nucleotide sequences • There are many different restriction enzymes that each cut DNA at different nucleotide sequences • Usually occurs at a palindrome 5’ GAATTC 3’ CTTAAG What is a palindrome in English? • Palindromes can be read backwards and forwards the same way. • Give me an example of a name that is a palindrome: – Hannah • It’s a little more tricky in DNA because the strand itself isn’t what must be backwards and forwards… DNA palindromes • A DNA palindrome reads exactly opposite its complementary strand • Ex- CAGAC is not a palindrome but CCCGGG is. Why? • Let’s look at the complementary strand: • CAGAC CCCGGG • GUCUG GGGCCC Another Example 17% 17% 17% 17% 17% 17% Which of these pieces of DNA represents a palindrome? 1. 5’ TTTCGC 3’ 3’ AAACCG 5’ 2. 5’ GAATTC 3’ 5’ CTTAAG 3’ 3. 5’ GAATTC 3’ 3’CTTAAG 5’ 4. 5’ AATGCC 3’ 5’ TTACGG 3’ Sticky Ends • Most restriction enzymes cut DNA with a staggered cut • The staggered cuts leave the DNA with end pieces “sticking off” – We call these “sticky ends” – These exposed N-bases will want to join with other complimentary exposed bases So what ? • What do you predict could happen if two pieces of DNA are cut with the same restriction enzyme??? – YES! They will have the same “sticky ends” – How could we use this??? • Restriction Enzymes can be used to make Recombinant DNA. • The gene of interest can be isolated using Restriction Enzymes Restriction Enzymes can be used to make RECOMBINANT DNA! • The gene you are interested in inserting (aka the “gene of interest”) can be cut using a restriction enzyme. • What will happen if I also cut the other organisms DNA with the same Restriciton Enzyme? Making Recombinant DNA… • Once the gene is isolated, how do we join it with the organism’s DNA? • Cut the organism’s DNA with the same restriction enzyme…why – The sticky ends will naturally be attracted to each other • Add DNA LIGASE: an enzyme that seals the fragments together Applications of this technology • PCR • GMO’s- (Transgenic Organisms) • Gene Therapy PCR • Now whenever the bacteria plasmid replicates, the gene will be cloned. • Gene Cloning: the production of many identical copies of the same gene. • You have created a Transgenic Organism! –organisms that contain functional recombinant DNA (rDNA) from a different organism Gene Therapy • Cystic Fibrosis example • http://learn.genetics.utah.edu/content/tech/ genetherapy/cysticfibrosis/ The Human Genome • A map of all the base pairs in human DNA. • Video- alopecia and the Human Genome Project The_Human_Genome_Project__Alopecia__and_the_Role_of_Genes_in_Disease.asf