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DNA TECHNOLOGY. O.J. Simpson capital murder case,1/95-9/95 Odds of blood in Ford Bronco not being R. Goldman’s: 6.5 billion to 1 Odds of blood on socks.

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Presentation on theme: "DNA TECHNOLOGY. O.J. Simpson capital murder case,1/95-9/95 Odds of blood in Ford Bronco not being R. Goldman’s: 6.5 billion to 1 Odds of blood on socks."— Presentation transcript:

1 DNA TECHNOLOGY

2 O.J. Simpson capital murder case,1/95-9/95 Odds of blood in Ford Bronco not being R. Goldman’s: 6.5 billion to 1 Odds of blood on socks in bedroom not being N. Brown-Simpson’s: 8.5 billion to 1 Odds of blood on glove not being from R. Goldman, N. Brown-Simpson, and O.J. Simpson: 21.5 billion to 1 Number of people on planet earth: 6.1 billion Odds of being struck by lightning in the U.S.: 2.8 million to 1 Odds of winning the Georgia’s Big Game lottery: 76 million to 1 Odds of getting killed driving to the gas station to buy a lottery ticket 4.5 million to 1 Odds of seeing 3 albino deer at the same time: 85 million to 1 Odds of having quintuplets: 85 million to 1 Odds of being struck by a meteorite: 10 trillion to 1

3 Biotechnology, defined broadly, is the engineering of organisms for useful purposes. Often, biotechnology involves the creation of hybrid genes and their introduction into organisms in which some or all of the gene is not normally present. Biotechnology Biotechnology, defined broadly, is the engineering of organisms for useful purposes. Often, biotechnology involves the creation of hybrid genes and their introduction into organisms in which some or all of the gene is not normally present. Recombinant DNA is DNA in which genes from 2 different sources are linked. Fourteen month-old genetically engineered (“biotech”) salmon (left) and standard salmon (right).

4 Manipulating Genes DNA Technology can be used to: DNA Technology can be used to: *cure diseases *to treat genetic disorders *to improve food crops *and to improve the lives of humans

5 Biotechnology Tools Restriction Enzymes- used to isolate a specific gene Restriction Enzymes- used to isolate a specific gene Cloning Vectors- a carrier that is used to clone a gene and transfer it from one organism to another. Example: Plasmid Cloning Vectors- a carrier that is used to clone a gene and transfer it from one organism to another. Example: Plasmid

6 Plasmids are Used to Replicate a Recombinant DNA *Plasmids are small circles of DNA found in bacteria. *Plasmids replicate independently of the bacterial chromosome. *Replication often produces copies of a recombinant plasmid in each cell. * Pieces of foreign DNA can be added within a plasmid to create a recombinant plasmid.

7 Restriction Enzymes are Enzymes That Cut DNA Only at Specific Sequences The enzyme EcoRI cutting DNA at its recognition sequence Different restriction enzymes have different recognition sequences. This makes it possible to create a wide variety of different gene fragments. Restriction enzyme animation

8 Forming Recombinant DNA using Restriction Enzymes:

9 Harnessing the Power of Recombinant DNA Technology – Human Insulin Production by Bacteria

10 Human Insulin Production by Bacteria 6) join the plasmid and human fragment and cut with a restriction enzyme

11 Human Insulin Production by Bacteria Mix the recombinant plasmid with bacteria.

12 Route to the Production by Bacteria of Human Insulin A fermentor used to grow recombinant bacteria. This is the step when gene cloning takes place. The single recombinant plasmid replicates within a cell. Then the single cell with many recombinant plasmids produces trillions of like cells with recombinant plasmid – and the human insulin gene. One cell with the recombinant plasmid

13 Route to the Production by Bacteria of Human Insulin The final steps are to collect the bacteria, break open the cells, and purify the insulin protein expressed from the recombinant human insulin gene.

14 Bacterial plasmids in gene cloning

15 DNA Technology Tools…. Polymerase Chain Reaction (DNA copying) Polymerase Chain Reaction (DNA copying) Gel Electrophoresis Gel Electrophoresis DNA Fingerprinting DNA Fingerprinting Cloning Cloning

16 Copying DNA Polymerase Chain Reaction Polymerase Chain Reaction Also called PCR Also called PCR A method of making many copies of a piece of DNA A method of making many copies of a piece of DNA

17 Steps in Copying DNA A DNA molecule is placed in a small test tube A DNA molecule is placed in a small test tube DNA polymerase that can work at high temps is added DNA polymerase that can work at high temps is added

18 Steps in Copying DNA The DNA is heated to separate the two strands The DNA is heated to separate the two strands Primers, short pieces of DNA complementary to the ends of the molecule to be copied, are added Primers, short pieces of DNA complementary to the ends of the molecule to be copied, are added

19 Copying DNA The tube is cooled, and DNA polymerase adds new bases to the separated strands The tube is cooled, and DNA polymerase adds new bases to the separated strands

20 PCR Large amounts of DNA can be made from a small starting sample

21 Electrophoresis DNA can be separated based on size and charge DNA can be separated based on size and charge The phosphate groups are negatively charged The phosphate groups are negatively charged DNA is placed in a gel and electricity is run through DNA is placed in a gel and electricity is run through

22 Electrophoresis Negative DNA moves toward the positive end Negative DNA moves toward the positive end Smaller fragments move farther and faster Smaller fragments move farther and faster

23 Electrophoresis

24 DNA Fingerprinting DNA is now a powerful tool in identification. Based on the fact that the amount of "junk DNA" differs uniquely between individuals. Structural genes are often separated by large regions of repeating basepairs. The number of these repeats is unique to an individual. Therefore when DNA from a person is cut with a restriction enzyme, the length of the fragments will be unique to an individual.

25 DNA Fingerprinting Continued… This will therefore produce a unique banding pattern following a gel electrophoresis. This will therefore produce a unique banding pattern following a gel electrophoresis. This test is highly accurate, and the probability of another individual possessing an identical banding pattern is estimated as around 1:14,000,000,000. This test is highly accurate, and the probability of another individual possessing an identical banding pattern is estimated as around 1:14,000,000,000.

26 DNA Fingerprinting

27 DNA Fingerprint EV shows the DNA at a crime scene. Samples 1-5 were taken and analyzed from suspects. Based on this evidence, which suspect is guilty?

28 Cloning Cloning is the process of making a genetically identical organism through nonsexual means. It has been used for many years to produce plants (even growing a plant from a cutting is a type of cloning). Animal cloning has been the subject of scientific experiments for years, but garnered little attention until the birth of the first cloned mammal in 1997, a sheep named Dolly. Since Dolly, several scientists have cloned other animals, including cows and mice. The recent success in cloning animals has sparked fierce debates among scientists, politicians and the general public about the use and morality of cloning plants, animals and possibly humans Cloning is the process of making a genetically identical organism through nonsexual means. It has been used for many years to produce plants (even growing a plant from a cutting is a type of cloning). Animal cloning has been the subject of scientific experiments for years, but garnered little attention until the birth of the first cloned mammal in 1997, a sheep named Dolly. Since Dolly, several scientists have cloned other animals, including cows and mice. The recent success in cloning animals has sparked fierce debates among scientists, politicians and the general public about the use and morality of cloning plants, animals and possibly humans Dolly, the first mammal clone

29 This shows how a common type of reproductive cloning works:

30 Dolly the Sheep Dolly died in This was due to lung disease and crippling arthritis. Most sheep can live years. Dolly was 6.5 years old.

31 Human Genome Project Started in 1990-finished in 2003 Started in 1990-finished in 2003 Research effort to sequence all of our DNA (46 chromosomes) Research effort to sequence all of our DNA (46 chromosomes) Over 3.3 billion nucleotides in roughly 26,000 genes Over 3.3 billion nucleotides in roughly 26,000 genes Mapping every gene location (loci) Mapping every gene location (loci) Conducted by scientists around the world Conducted by scientists around the world

32 HGP Insights…what they learned Only 2% of human genome codes for proteins (exons) Only 2% of human genome codes for proteins (exons) Other 98% (introns) are non- coding Other 98% (introns) are non- coding Only about 26,000 genes (expected 100,000) Only about 26,000 genes (expected 100,000) Proteome – organism’s complete set of proteins Proteome – organism’s complete set of proteins About 8 million single nucleotide polymorphisms (SNP) – places where humans differ by a single nucleotide About 8 million single nucleotide polymorphisms (SNP) – places where humans differ by a single nucleotide

33 Benefits of Human Genome Project Improvements in medical prevention of disease, gene therapies, diagnosis techniques … Improvements in medical prevention of disease, gene therapies, diagnosis techniques … Production of useful protein products for use in medicine, agriculture, bioremediation and pharmaceutical industries. Production of useful protein products for use in medicine, agriculture, bioremediation and pharmaceutical industries. Improved bioinformatics – using computers to help in DNA sequencing … Improved bioinformatics – using computers to help in DNA sequencing …

34 Gene Therapy Gene therapy can be used to treat some genetic disorders. It is utilized when a a defective “unhealthy” gene is replaced with normal “healthy” gene. Here are some diseases that gene therapy may help with more research: Cystic fibrosis Cystic fibrosis Hemophilia Hemophilia Lung Cancer Lung Cancer AIDS AIDS Ovarian Cancer Ovarian Cancer Brain tumors Brain tumors

35 Example of gene therapy 1. Insert DNA version of normal allele into virus. 2. Let virus infect bone marrow cells that have been removed from the patient and cultured. 3. Viral DNA carrying the normal allele inserted into chromosome. Insert a gene into an inactivated or nonvirulent virus and use the virus's infective capabilities to carry the desired gene into the patient's cells.

36 What factors have kept gene therapy from becoming an effective treatment for genetic disease? Immune response Problems with viral vectors Multigene disorders

37 BiotechnologyBiotechnology The use of gene science to create new products from plants and animals The use of gene science to create new products from plants and animals Biotechnology Provides Biotechnology Provides : Improved food products Medical advances

38 Biotechnology Breakthroughs…. Insulin (1982) Insulin (1982) First commercial biotech product First commercial biotech product Reliable, inexpensive source of insulin Reliable, inexpensive source of insulin Rice Rice Enriched with beta-carotene and iron Enriched with beta-carotene and iron Bananas Bananas Containing edible hepatitis vaccine Containing edible hepatitis vaccine Potatoes with higher solid content Potatoes with higher solid content Garlic that lowers cholesterol Garlic that lowers cholesterol Fruits and vegetables that reducerisks of cancer and heart disease Fruits and vegetables that reducerisks of cancer and heart disease

39 Herbicide Resistant Crops Soybeans: Roundup Ready Soybeans: Roundup Ready Corn: Roundup Ready, Liberty Link Corn: Roundup Ready, Liberty Link Cotton: BXN, Roundup Ready Cotton: BXN, Roundup Ready Canola: Liberty Link, Roundup Ready Canola: Liberty Link, Roundup Ready + CP4 EPSPS = Roundup gene Ready


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