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Chapter 13 - Biotechnology

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1 Chapter 13 - Biotechnology
Biology Metz Chapter 13 - Biotechnology What is biotechnology? What is biotechnology used for? When did biotechnology begin? What are the basic techniques used in modern biotechnology? How is biotechnology influencing human medicine? Chapter 13 Chapter 13

2 What is biotechnology? Alteration of organisms, cells, or biological molecules to achieve specific practical goals Chapter 13

3 What is biotechnology used for?
Production of pharmaceuticals Production of commercial compounds Agricultural improvements Environmental cleanup To better understand the functions and interactions of genes -Advances in medical treatments -Advances in agriculture Diagnosis of genetic disorders Forensics Chapter 13

4 When did biotechnology begin?
Genetic manipulation/alteration of organisms has been occurring for over 10,000 years Chapter 13

5 Historical vs. modern genetic manipulation
Historical – breeding Chapter 13

6 Domestication of crops began ~10,000B.C.
Chapter 13

7 Genetic manipulation in the Brassica family
Europe & Mediterranean Chapter 13

8 Teosinte – ancestor of corn (Mexico)
Cob selection 5000BC – 2cm 3000BC – 4cm 1000AD – 13cm Chapter 13

9 What are the basic techniques of modern biotechnology?
Recombinant DNA technology -Moving small pieces of DNA between organisms -Cloning Genes -Isolating DNA from cells -Restriction enzymes -Polymerase Chain Reaction -Gel electrophoresis -DNA probes -Genetic engineering Viral mediated gene transfer Gene Gun Agrobacterium tumefaciens DNA microarrays Chapter 13

10 Recombinant DNA technology
Combining genes or parts of genes from different organisms Transgenic – organisms containing “foreign” genes Genetically modified organisms = GMOs Chapter 13

11 Recombinant products rBST and dairy products Chapter 13

12 DNA recombination occurs naturally
Is combining genes from different species inappropriately tampering with God’s creation? DNA recombination occurs naturally 1. Sexual reproduction 2. Bacterial transformation 3. Viral transfer of DNA Chapter 13

13 Technologies used in cloning genes
Isolating DNA from cells Restriction enzymes Gel electrophoresis Polymerase chain reaction Hybridization Chapter 13

14 Isolating DNA from cells
Cell membranes are lysed in detergent Proteins are removed by precipitation DNA is precipitated with alcohol Chapter 13

15 Restriction enzymes cut DNA molecules
Restriction endonucleases (enzymes) cut DNA at specific short palindromic nucleotide sequences Chapter 13

16 Agarose gel electrophoresis
Chapter 13

17 Gel electrophoresis separates DNA fragments by size
Chapter 13

18 Polymerase chain reaction (PCR)
Copies specific DNA sequences A thermal cycler Chapter 13

19 Polymerase chain reaction (PCR)
25-30 PCR cycles yield ~1 million copies of the DNA sample Can be used on trace quantities of DNA Saliva, hair follicle, trace dried blood Chapter 13

20 Hybridization – detection of specific DNA sequences (utilizes complementary base pairing)
Southern blotting animation Chapter 13

21 Forensic DNA analysis & hybridization
RFLP (Restriction Fragment Length Polymorphism) analysis Began in 1985 Chapter 13

22 RFLP analysis and hybridization
Chapter 13

23 RFLP Case Study - OJ Simpson
Chapter 13

24 Chapter 13

25 Hybridization & medicine: Diagnosis
RFLP analysis can be used in genetic disease diagnosis (sickle-cell anemia) Chapter 13

26 Modern forensic DNA analysis
Based on the Polymerase Chain Reaction and STRs (Short tandem repeats) Chapter 13

27 STRs and identity Number of repeats varies (i.e. There are numerous STR “alleles”) Chapter 13

28 PCR, STRs and identity The greater the number of repeats, the longer the DNA pieces amplified by PCR Chapter 13

29 What are the basic techniques of modern biotechnology?
Genetic engineering - Addition, deletion, or modification of genes in an organism Genetic engineering is not limited to the genome of the organism being manipulated Chapter 13

30 Genetic engineering & agriculture
Chapter 13

31 Genetic engineering in plants: Gene gun
Biology Metz Genetic engineering in plants: Gene gun Figure: FIGURE 13.7 Title: Inserting genes into plant cells Caption: Genetic engineering techniques have been used to insert a firefly gene that codes for the enzyme luciferase into this tobacco plant. The enzyme breaks down the chemical luciferin, releasing light in the process. This plant has taken up water containing luciferin. Chapter 13 Chapter 13

32 Genetic engineering in plants: Agrobacterium tumefaciens
Biology Metz Genetic engineering in plants: Agrobacterium tumefaciens Figure: FIGURE 13.E1b Title: Biotechnology may help the American chestnut Caption: Chestnut blight fungus has produced a lesion on this chestnut tree. The lesioned area was treated with infected fungus, and the tree is walling off the infection with a growth of new tissue. Chapter 13 Chapter 13

33 Plant biotechnology: Insect resistant crop plants
Bacillus thuringiensis (BT) Chapter 13

34 Bacillus thuringiensis
BT spores form insecticidal crystal proteins Chapter 13

35 Plant biotechnology: Herbicide resistant plants
Chapter 13

36 Genetic engineering of animals
Genzyme GFP Salmon growth hormone gene engineered for constitutive (always on) expression Chapter 13

37 How is biotechnology influencing human medicine?
Disease diagnosis and DNA arrays Microarray construction link Chapter 13

38 How is biotechnology influencing human medicine?
Correlating genes & disease Chapter 13

39 Biotechnology & whole genome analysis
Whole genome arrays ~$ each Chapter 13

40 How is biotechnology influencing human medicine?
Chapter 13

41 Somatic Cell Gene Therapy
Biology Metz Somatic Cell Gene Therapy Figure: FIGURE 13.11 Title: Hope through gene therapy Caption: (a) Andrew Gobea (shown with his mother) received gene therapy on stem cells as a newborn. Eventually, he may produce adequate numbers of normal immune cells on his own. (b) Genetic engineering of stem cells may permanently replace the defective ADA gene. 1) Stem cells are harvested from umbilical cord blood. 2) A retrovirus engineered to contain normal human ADA genes is mixed with the stem cells in culture. 3) The retrovirus transmits its DNA, including the functional gene, into the stem cells. 4) The engineered stem cells are injected into the same newborn to take up residence in bone marrow and produce normal immune cells. Chapter 13 Chapter 13

42 Germline Gene Therapy Chapter 13 Biology 111 - Metz Chapter 13
Figure: FIGURE 13.13 Title: Human cloning technology might allow permanent correction of genetic defects Caption: Researchers might derive human embryos from eggs fertilized in culture dishes, using sperm and eggs from the natural parents, one or both of whom have a genetic disorder. When an embryo containing a defective gene grows into a small cluster of cells, a single cell could be removed from the embryo and the defective gene replaced by means of an appropriate vector. Then the repaired nucleus could be implanted into another egg (taken from the mother) whose nucleus had been removed. The repaired, diploid egg cell could then be implanted in the mother’s uterus for normal development. Chapter 13 Chapter 13

43 American Journal of Medicine, Nov. 2003, p. 563
Chapter 13 American Journal of Medicine, Nov. 2003, p. 563

44 Biotechnology is regulated
Chapter 13

45 Biotechnology concerns - allergenicity
Chapter 13

46 Biotechnology concerns
Gene “escape” via pollen Chapter 13

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