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Points to Ponder• What are three functions of DNA?• Review DNA and RNA structure. • What are the 3 types of RNA and what are their functions?• Compare and contrast the structure and function of DNA and
RNA.• How does DNA replicate?• Describe transcription and translation in detail.• Describe the genetic code.• Review protein structure and function.• What are the 4 levels of regulating gene expression.• What did we learn from the human genome project and where
are we going from here?• What is ex vivo and in vivo gene therapy?• Define biotechnology, transgenic organisms, genetic
engineering and recombinant DNA.• What are some uses of transgenic bacteria, plants and
animals?
What must DNA do?
1. Replicate to be passed on to the next generation
2. Store information
3. Undergo mutations to provide genetic diversity
21.1 DNA and RNA structure and function
DNA structure: A review• Double-stranded helix
• Composed of repeating nucleotides (made of a pentose sugar, phosphate and a nitrogenous base)
• Sugar and phosphate make up the backbone while the bases make up the “rungs” of the ladder
• Bases have complementary pairing with cytosine (C) pairs with guanine (G) and adenine (A) pairs with thymine (T)
21.1 DNA and RNA structure and function
DNA structure
21.1 DNA and RNA structure and function
How does DNA replicate?
• The two strands unwind by breaking the H bonds
• Complementary nucleotides are added to each strand by DNA polymerase
• Each new double-stranded helix is made of one new strand and one old strand (semiconservative replication)
• The sequence of bases makes each individual unique
21.1 DNA and RNA structure and function
DNA replication
21.1 DNA and RNA structure and function
RNA structure and function• Single-stranded
• Composed of repeating nucleotides
• Sugar-phosphate backbone
• Bases are A, C, G and uracil (U)
• Three types of RNA– Ribosomal (rRNA): joins with proteins to form ribosomes– Messenger (mRNA): carries genetic information from DNA to
the ribosomes– Transfer (tRNA): transfers amino acids to a ribosome where
they are added to a forming protein
21.1 DNA and RNA structure and function
RNA structure21.1 DNA and RNA structure and function
Comparing DNA and RNA
• Similarities: – Are nucleic acids– Are made of nucleotides– Have sugar-phosphate backbones– Are found in the nucleus
• Differences:– DNA is double
stranded while RNA is single stranded
– DNA has T while RNA has U
– RNA is also found in the cytoplasm as well as the nucleus while DNA is not
21.1 DNA and RNA structure and function
Proteins: A review• Composed of subunits of amino acids
• Sequence of amino acids determines the shape of the protein
• Synthesized at the ribosomes
• Important for diverse functions in the body including hormones, enzymes and transport
• Can denature causing a loss of function
21.2 Gene expression
2 steps of gene expression
1. Transcription – DNA is read to make a mRNA in the nucleus of our cells
2. Translation – Reading the mRNA to make a protein in the cytoplasm
21.2 Gene expression
Overview of transcription and translation
21.2 Gene expression
The genetic code
• Made of 4 bases
• Bases act as a code for amino acids in translation
• Every 3 bases on the mRNA is called a codon that codes for a particular amino acid in translation
21.2 Gene expression
1. Transcription
• mRNA is made from a DNA template
• mRNA is processed before leaving the nucleus
• mRNA moves to the ribosomes to be read
• Every 3 bases on the mRNA is called a codon and codes for a particular amino acid in translation
21.2 Gene expression
Processing of mRNA after transcription
Modifications of mRNA:• One end of the RNA is capped
• Introns removed
• Poly-A tail is added
21.2 Gene expression
Visualizing the 3 steps of translation21.2 Gene expression
How can we modify a person’s genome?
• Gene therapy - insertion of genetic material into human cells to treat a disorder
– Ex vivo therapy – cells are removed for a person altered and then returned to the patient
– In vivo therapy – a gene is directly inserted into an individual through a vector (e.g. viruses) or directly injected to replace mutated genes or to restore normal controls over gene activity
• Gene therapy has been most successful in treating cancer
21.3 Genomics
Ex vivo gene therapy
21.3 Genomics
DNA technology terms
• Genetic engineering – altering DNA in bacteria, viruses, plants and animal cells through recombinant DNA techonology
• Recombinant DNA – contains DNA from 2 or more different sources
• Transgenic organisms – organisms that have a foreign gene inserted into them
• Biotechnology – using natural biological systems to create a product or to achieve an end desired by humans
21.4 DNA technology
DNA technology
1. Gene cloning through recombinant DNA
2. Polymerase chain reaction (PCR)
3. DNA fingerprinting
4. Biotechnology products from bacteria, plants and animals
21.4 DNA technology
1. Gene cloning
• Recombinant DNA – contains DNA from 2 or more different sources that allows genes to be copies
• An example using bacteria to clone the human insulin gene:
– Restriction enzyme – used to cut the vector (plasmid) and the human DNA with the insulin gene
– DNA ligase: seals together the insulin gene and the plasmid
– Bacterial cells uptake plasmid and the gene is copied and product can be made
21.4 DNA technology
Visualizing gene cloning21.4 DNA technology
2. Polymerase chain reaction (PCR)
• Used to clone small pieces of DNA• Important for amplifying DNA for analysis
such as in DNA fingerprinting
21.4 DNA technology
3. DNA fingerprinting
• Fragments are separated by their charge/size ratios• Results in a distinctive pattern for each individual• Often used for paternity or to identify an individual at a
crime scene or unknown body remains
21.4 DNA technology
4. Biotechnology products: Transgenic bacteria
• Important uses:– Insulin– Human growth
hormone (HGH)– Clotting factor VIII– Tissue plasminogen activator (t-PA)– Hepatitis B vaccine– Bioremediation – cleaning up the
environment such as oil degradingbacteria
21.4 DNA technology
4. Biotechnology products: Transgenic plants
• Important uses:– Produce human proteins in their seeds such as
hormones, clotting factors and antibodies– Plants resistant to herbicides– Plants resistant to insects– Plants resistant to frost
• Corn, soybean and cotton plants are commonly genetically altered
• In 2001:– 72 million acres of transgenic crops worldwide– 26% of US corn crops were transgenic crops
21.4 DNA technology
4. Biotechnology products: Transgenic plants
21.4 DNA technology
Health focus: Ecological concern about BT crops
• Resistance increasing in the target pest
• Exchange of genetic material between the transgenic plant and a related species
• Concern about the impact of BT crops on nontarget species
21.4 DNA technology
4. Biotechnology products: Transgenic animals
• Gene is inserted into the egg that when fertilized will develop into a transgenic animal
• Current uses:– Gene pharming: production of pharmaceuticals in the
milk of farm animals– Larger animals: includes fish, cows, pigs, rabbits and
sheep– Mouse models: the use of mice for various gene
studies– Xenotransplantation: pigs can express human
proteins on their organs making it easier to transplant them into humans
21.4 DNA technology