Chapter 7

1. Chapter 7 Genetically Modified OrganismsGene Expression, Mutation, and Cloning 0

2. 1 Protein Synthesis and Gene Expression In the early 1980s, genetic engineers began producing recombinant bovine growth hormone (rBGH) Made by genetically engineered bacteria The bacteria were given DNA that carries instructions for making BGH In cows, growth hormones increase body size and milk production

3. 1 Protein Synthesis and Gene Expression: From Gene to Protein Protein synthesis – the process of using instructions carried on a gene to create proteins. Several steps are involved and require both DNA and RNA. Gene –a sequence of DNA that encodes a protein Protein – a large molecule composed of amino acids

4. 1 Protein Synthesis and Gene Expression: From Gene to Protein DNA Double-stranded Each nucleotide composed of deoxyribose, phosphate, and nitrogenous base 4 bases: adenine, thymine, guanine, cytosine

5. 1 Protein Synthesis and Gene Expression: From Gene to Protein RNA Single-stranded Nucleotides comprised of ribose, phosphate, and nitrogenous base 4 bases: A, T, G, and Uracil

6. 1 Protein Synthesis and Gene Expression: From Gene to Protein The flow of genetic information in a cell is DNARNA protein and occurs in 2 steps: Transcription (DNA  RNA) Translation (RNA  Protein)

7. 1 Protein Synthesis and Gene Expression: Transcription Transcription occurs in the nucleus. RNA polymerase binds to the promoter region of the gene. RNA polymerase zips down the length of gene, matching RNA nucleotides with complementary DNA nucleotides This forms messenger RNA (mRNA)

8. Animation: Transcription Click “Go to Animation” / Click “Play”

9. 1 Protein Synthesis and Gene Expression: Translation Translation occurs in the cytoplasm (outside the nucleus). Translation requires: mRNA (made during transcription), amino acids, energy (ATP), and some helper molecules. Ribosomes Transfer RNA (tRNA)

10. 1 Protein Synthesis and Gene Expression: Translation Ribosomes The ribosome is composed of ribosomal RNA (rRNA) and comprises a small and a large subunit.

11. 1 Protein Synthesis and Gene Expression: Translation Transfer RNA: tRNA carries amino acids and matches its anticodon with codons on mRNA Codons are 3 nucleotides long

12. 1 Protein Synthesis and Gene Expression: Translation A protein is put together one amino acid at a time. The ribosome attaches to the mRNA at the promoter region. Ribosome facilitates the docking of tRNA anticodons to mRNA codons. When two tRNAs are adjacent, a bond is formed between their amino acids. Forms a peptide chain of amino acid

13. 1 Protein Synthesis and Gene Expression: Translation

14. 1 Protein Synthesis and Gene Expression: Translation

15. 1 Protein Synthesis and Gene Expression: Genetic Code The genetic code allows a specific codon to code for a specific amino acid. A codon is comprised of three nucleotides = 64 possible combinations (43 combinations) 61 codons code for amino acids 3 others are stop codons, which end protein synthesis Genetic code expresses redundancy The genetic code is universal

16. 1 Protein Synthesis and Gene Expression: Genetic Code

17. Animation: Translation Click “Go to Animation” / Click “Play” BioFlix: Protein Synthesis

18. 1 Protein Synthesis and Gene Expression: Mutations Changes in genetic sequence = mutations Changes in genetic sequence might affect the order of amino acids in a protein. Protein function is dependent on the precise order of amino acids Possible outcomes of mutation: 1 - no change in protein 2 - non-functional protein 3 - different protein

19. 1 Protein Synthesis and Gene Expression: Mutation Base-substitution mutation Simple substitution of one base for another

20. 1 Protein Synthesis and Gene Expression: Mutation Neutral mutation Mutation does not change the function of the protein, it codes for the same amino acid

21. 1 Protein Synthesis and Gene Expression: Mutation Frameshift mutation Addition or deletion of a base, which changes the reading frame

22. 1 Protein Synthesis and Gene Expression: An Overview of Gene Expression Each cell in your body (except sperm and egg cells) has the same DNA. But each cell only expresses a small percentage of genes. Example: Nerve and muscle cells perform very different functions, thus they use different genes. Turning a gene or a set of genes on or off = regulating gene expression

23. 1 Protein Synthesis and Gene Expression: An Overview of Gene Expression Nerves and cells have the same suite of genes, but they express different genes.

24. 1 Protein Synthesis and Gene Expression: Regulating Gene Expression Regulation of transcription Prokaryotic cells use repressors to regulate gene expression Repressors bind to the promoter and prevent the RNA polymerase from binding

25. 1 Protein Synthesis and Gene Expression: Regulating Gene Expression Regulation of transcription Eukaryotic cells use activators to regulate gene expression Activators help the RNA polymerase bind to the promoter

26. 1 Protein Synthesis and Gene Expression: Regulating Gene Expression Regulation by chromosome condensation Folding up of the chromosomes prevents transcription Regulation by mRNA degradation Nucleases cut mRNA Regulation of Translation Slowing of binding of the mRNA to the ribosome Regulation of Protein Degradation Proteases degrade proteins

27. 2 Producing Recombinant Proteins: Cloning a Gene Using Bacteria rBGH is a protein, and is coded by a specific gene. Transfer of rBGH gene to bacteria allows for growth under ideal conditions. Bacteria can serve as “factories” for production of rBGH. Cloning of the gene is making many copies of that gene.

28. 2 Producing Recombinant Proteins: Cloning a Gene Using Bacteria Restriction enzymes – Used by bacteria as a form of defense. Restriction enzymes cut DNA at specific sequences. They are important in biotechnology because they allow scientists to make precise cuts in DNA. Plasmid – Small, circular piece of bacterial DNA that exists separate from the bacterial chromosome. Plasmids are important because they can act as a ferry to carry a gene into a cell.

29. 2 Producing Recombinant Proteins: Cloning a Gene Using Bacteria Step 1. Remove the gene from the cow chromosome

30. 2 Producing Recombinant Proteins: Cloning a Gene Using Bacteria Step 2. Insert the BGH gene into the bacterial plasmid

31. 2 Producing Recombinant Proteins: Cloning a Gene Using Bacteria Recombinant – Indicates material that has been genetically engineered: a gene that has been removed from its original genome and combined with another. After step 2, the GBH is now referred to as recombinant GBH or rGBH.

32. 2 Producing Recombinant Proteins: Cloning a Gene Using Bacteria Step 3. Insert the recombinant plasmid into a bacterial cell

33. 2 Producing Recombinant Proteins: Cloning a Gene Using Bacteria About 1/3 of cows in the US are injected with rBGH. rBGH increases milk volume from cows by about 20%. The same principles apply to other proteins. Clotting proteins for hemophiliacs are produced using similar methods. Insulin for diabetics is also produced in this way. FDA approval is needed for any new food that is not generally recognized as safe (GRAS).

34. Animation: Producing Bovine Growth Hormone Click “Go to Animation” / Click “Play”

36. 3 Genetically Modified Foods All agricultural products are the result of genetic modification through selective breeding. Artificial selection does not move genes from one organism to another, but does drastically change the characteristics of a population. Genetically modifying foods Increase shelf life, yield, or nutritional value Golden rice has been genetically engineered to produce beta-carotene, which increases the rice’s nutritional yield.

37. 3 Genetically Modified Foods: Modifying Plants with the Ti Plasmid and Gene Gun Unlike rBGH, crop plants are directly modified. In order to do this, the target gene must be inserted into the plant cell. Two methods to do this: Ti plasmid Gene gun

38. 3 Genetically Modified Foods: Modifying Plants with the Ti Plasmid

39. 3 Genetically Modified Foods: Modifying Plants with the Gene Gun

40. 3 Genetically Modified Foods: Modifying Plants with the Ti Plasmid and Gene Gun Transgenic organism – the result of the incorporation of a gene from one organism to the genome of another. Also referred to as a genetically modified organism (GMO). Benefits: Crops can be engineered for resistance to pests, thus farmers can spray fewer chemicals. Concerns: Pests can become resistant to chemicals. GM crops may actually lead to increased use of pesticides and herbicides. GM crop plants may transfer genes to wild relatives.

41. 4 Genetically Modified Humans: Stem Cells Stem cells – undifferentiated cells, capable of growing in to many different kinds of cells and tissues Stems cells might be used to treat degenerative diseases such as Alzheimer’s or Parkinson’s. Using stem cells to produce healthy tissue is called therapeutic cloning. Stem cells could also be used to grow specific tissues to treat burns, heart attack damage, or replacement cartilage in joints. Stems cells are totipotent, meaning they can become any other cell in the body.

42. 4 Genetically Modified Humans: Human Genome Project Human Genome Project – international effort to map the sequence of the entire human genome (~20,000 – 25,000 genes). For comparative purposes, genomes of other model organisms (E. coli, yeast, fruit flies, mice) were also mapped. It was sequenced using the technique of chromosome walking.

43. 4 Genetically Modified Humans: Gene Therapy Gene therapy – replacement of defective genes with functional genes Germ line gene therapy Embryonic treatment Embryo supplied with a functional version of the defective gene. Embryo + cells produced by cell division have a functional version of gene. Somatic cell gene therapy Somatic cell gene therapy – fix or replace the defective protein only in specific cells

44. 4 Genetically Modified Humans: Gene Therapy Somatic cell therapy used as a treatment of SCID (severe combined immunodeficiency) All somatic cells have limited lifetimes. Therapy is not permanent and requires several treatments per year.

45. 4 Genetically Modified Humans: Cloning Humans Human cloning occurs naturally whenever identical twins are produced. Cloning of offspring from adults has already been done with cattle, goats, mice, cats, pigs, and sheep. Cloning is achieved through the process of nuclear transfer.

46. 4 Genetically Modified Humans: Cloning Humans

47. Which of the following types of RNA carries amino acids to the growing polypeptide chain? mRNA tRNA rRNA RNA does not carry amino acids

48. Which of the following types of RNA carries amino acids to the growing polypeptide chain? mRNA tRNA rRNA RNA does not carry amino acids

49. A sequence of mRNA, called a codon, reads ACU. How will the set of nucleotides on the anticodon of the tRNA read? ACU UGA TGA AUG

50. A sequence of mRNA, called a codon, reads ACU. How will the set of nucleotides on the anticodon of the tRNA read? ACU UGA TGA AUG