1. Biogenetic Engineering & Manipulating Genes
Chapter 20
Genomics video: Annenberg series

2. Intro. Q’s #5 for Chapter 20: Genetic Engineering
What does the acronym PCR stand for and what does this process do?
What does Gel electrophoresis allow us to do?
Give two applications of DNA profiling.
What are the advantages and disadvantages of genetic screening?
How is a cDNA library different from a genomic library? (p )
Name two “vectors” that can be used for gene transfer.
Give two examples of a genetically modified crop or animal.
Briefly explain the process of gene therapy and give an example how it works.
Explain what a clone is and how it could be formed.
What are some of the ethical concerns about cloning? Give your opinion if you think cloning is something we should be doing.

3. Genetic Engineering Chapter 20 DNA Technology & Genomics
Pgs &

4. 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 Illinois 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:
Odds of being struck by a meteorite:
10 trillion to 1

5. Introduction
Manipulating genes and sequencing DNA has become one of the greatest achievements in the last 50 years -First complete genome sequenced was a bacteria (1995)
-Human Genome project ( )

6. Recombinant DNA
Def: DNA in which genes from 2 different sources are linked
Genetic engineering: direct manipulation of genes for practical purposes
Biotechnology: manipulation of organisms or their components to perform practical tasks or provide useful products

7. Tools for DNA Analysis & Genomics
PCR (polymerase chain reaction)
Gel electrophoresis
Restriction fragment analysis (RFLPs)
Southern blotting
DNA sequencing
Human genome project

8. Tools of Genetic Engineering
Restriction enzymes (endonucleases)
-in nature, these enzymes protect bacteria from intruding DNA; they cut up the DNA (restriction); very specific
Restriction site:
-recognition sequence for a particular restriction enzyme
Restriction fragments:
-segments of DNA cut by restriction enzymes in a reproducable way
Sticky end:
-short extensions of restriction fragments
DNA ligase:
-enzyme that can join the sticky ends of DNA fragments
Cloning vector:
-DNA molecule that can carry foreign DNA into a cell and replicate there (usually bacterial plasmids)

9. Restriction Enzymes (endonucleases) http://highered. mcgraw-hill

10. Producing Restriction Fragments
DNA ligase used to splice together cut plasmids and chromosome fragments

11. Cloning

12. Steps for Eukaryotic Gene Cloning (pg. 387)
Isolation of cloning vector (bacterial plasmid) & gene-source DNA (gene of interest)
Insertion of gene-source DNA into the cloning vector using the same restriction enzyme; bind the fragmented DNA with DNA ligase
Introduction of cloning vector into cells (transformation by bacterial cells)
Cloning of cells (and foreign genes)
Identification of cell clones carrying the gene of interest

13. Steps for Eukaryotic gene cloning13

14. DNA Libraries (pg. 388)
Collection of DNA fragments that have been incorporated into plasmids

15. Genomic Libraries (pg. 388)
Stored amount of cloned genes within bacteria or virues (bacteriophages)
Contained as cloned plasmids within bacteria or as fragments within Bacteriophages genome
Phages can carry a larger DNA insert vs. bacteria

16. cDNA Library (Pg. 390) cDNA (complementary DNA)
Start w/mRNA to make a ssDNA strand
Use an enzyme “reverse transcriptase” which is found in retroviruses such as HIV (see pg ) to make a ssDNA strand.
A complementary strand of DNA is made by DNA polymerase to produce a dsDNA molecule (cDNA)

17. Polymerase Chain Reaction (PCR) http://highered. mcgraw-hill
Amplification of any piece of DNA without cells (in vitro)
Produces many identical copies of a DNA segment
Materials: heat, DNA polymerase, nucleotides, single-stranded DNA primers
Applications: fossils, forensics, prenatal diagnosis, etc.

18. Polymerase Chain Reaction
PCR = common method of creating copies of specific fragments of DNA
PCR rapidly amplifies a single DNA molecule into many billions of molecules.
Small samples of DNA can produce sufficient copies to carry out forensic tests

19. Polymerase Chain reaction (PCR)

20. Inserting DNA into the nucleus of a cell

21. Gel Electrophoresis (Lab)
DNA fragments placed into “wells” in gel agarose
Electricity pulls on DNA fragments
Fragments travel at different rates based on size and ability to squeeze through swiss-cheese-like agarose

22. Restriction Fragment Analysis
Restriction fragment length polymorphisms (RFLPs)
Southern blotting: process that reveals sequences and the RFLPs in a DNA sequence
DNA Fingerprinting (DNA Profiling)

23. Gel Electrophoresis
separates nucleic acids or proteins on the basis of size and electrical charge creating DNA bands of the same length
DNA has a net negative charge (use a positive charge in the gel)

24. Intro. Q’s #5 for Chapter 20: Genetic Engineering
What does the acronym PCR stand for and what does this process do?
What does Gel electrophoresis allow us to do?
Give two applications of DNA profiling.
What are the advantages and disadvantages of genetic screening?
How is a cDNA library different from a genomic library? (p )
Name two “vectors” that can be used for gene transfer.
Give two examples of a genetically modified crop or animal.
Briefly explain the process of gene therapy and give an example how it works.
Explain what a clone is and how it could be formed.
What are some of the ethical concerns about cloning? Give your opinion if you think cloning is something we should be doing. 24

25. Video #3: Using DNA evidence to Solve Crimes
Write 15 key statements from the video.
Please number each statement.
Indicate where the sources of DNA were found for each case.

26. 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 Illinois 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:
Odds of being struck by a meteorite:
10 trillion to 1 26

27. DNA Profiling (DNA fingerprinting)
Applications:
-Used in criminal investigations
-Identify the remains of dead people
-Compare Evolutionary origins of organisms
-Look for genetic disorders (mutations)
-Sequence DNA samples

28. DNA Fingerprinting

29. Southern Blot of DNA Fragments (pg. 395)
Used to determine the presence or absence of a particular nucleotide sequence in the DNA different sources.
Compare the restriction fragments produced from the different samples.
Radioactive probes are used and attach to the DNA fragments by complementary base pairing.

30. Practical DNA Technology Uses
Diagnosis of disease
Human gene therapy
Pharmaceutical products (vaccines)
Forensics
Animal husbandry (transgenic organisms)
Genetic engineering in plants
Ethical concerns?

31. Genetic Screening
Def: Testing individuals in a population for the presence or absence of a gene (allele)
Advantages:
-pre-natal diagnosis of genetic disorders
-Could help stop the spread of a disorder
-Can detect carriers of a potential disorder
Disadvantages:
-invasion of privacy
-Individuals can become stigmatized in the community
-Discriminated against or feared
-Employment and medical insurance

32. DNA Sequencing
Determination of nucleotide sequences (Sanger method, sequencing machine)
Genomics: the study of genomes based on DNA sequences
Human Genome Project

33. Bacterial plasmids in gene cloning

34. Plant Cloning Tissue Culture Propagation
Bits of phloem can be induced in the lab to form clumps of tissue that will make roots & shoots
Orchid culture

35. Embryo Cloning
Medical technique which produces identical twins or triplets
Duplicates nature
One or more cells are removed from a fertilized embryo, encouraged to develop into one identical twins or triplets
Done for many years on animals
Limited experimentation on humans

36. Adult DNA Cloning Cell nuclear replacement
Produces a duplicate of an existing animal
DNA from an embryo is removed;
Replaced with DNA from an adult animal
Embryo is implanted in a womb and allowed to develop into a new animal
Untried on humans - potential of producing a twin of an existing person

37. Therapeutic Cloning
Stem cells removed from an embryo with intent of producing tissue or a whole organ for transplant back into the person who supplied the new DNA
Embryo dies in the process
Goal is to produce a healthy copy of a sick person's tissue or organ for transplant

38. Application: Gene Therapy38

39. Therapeutic Cloning Vastly superior to organ transplants
Supply would be unlimited - no waiting lists
Tissue or organ would have the sick person's original DNA
No immunosuppressant drugs would need to be taken

40. Adult DNA Cloning

41. Adult DNA Cloning

42. Stem
cell
clon-ing

44. Theraputic Stem Cell Cloning
Used

45. Chromatin Def: complex of DNA and proteins DNA Packing
Histone proteins
(+ charged amino acids w/ phosphates of DNA that are - charged)
Nucleosome
-”beads on a string”; basic unit of DNA packing
Heterochromatin
-highly condensed interphase DNA (can not be transcribed)
Euchromatin
-less compacted interphase DNA (can be transcribed)