Ms.Sastry, AP Biology
Unit 3/Chapter 20
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Chapter 20 – Biotechnology follow along in class notes
Free response questions/objectives of chapter:
1) Describe how recombinant DNA may be used to clone human genes.
2) What is PCR – how is it use in Biotechnology?
3) What is DNA gel electrophoresis – what are its principles? How is used to analyze RFLPs and what
are its applications?
4) What is nucleic acid hybridization? How is it used in Biotechnology?
1) What is biotechnology? Give an example of a product of this industry.
2) How is the biotechnology industry different from the pharmaceutical industry?
3) What is recombinant DNA? Which industry will it be used in?
4) What are the goals of Biotechnology?
Goal1: Isolate gene of interest: how are restriction
enzymes used for this process?
Goal 2: Make recombinant DNA containing gene of
interest
Ms.Sastry, AP Biology
Unit 3/Chapter 20
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Biotechnology goals continued:
Goal 3: Clone the recombinant DNA containing gene of interest using a vector
Ex: 1 – clone in bacteria using plasmid vector – what does this mean?
Steps: 1) Insert recombinant plasmid into bacteria by BACTERIAL TRANSFORMATION
Goal 3: Ex 2: clone in yeast (eukaryotic cells) – How is this different? What are some problems with human
(eukaryotic) gene cloning?
Step 2a: Clonal screening using antibiotics and markers/reporter gene: what is the purpose of clonal screening and
how is it done?
Draw recombinant
plasmid here:
Goal 3: Step 2b: Clonal screening and selection using
radioactive/fluorescent probes: nucleic acid hybridization
Goal 3: continued on a larger scale – DNA not just from a single gene of interest, but many, many such genes or in
fact ALL the human genes could be stored in a:
Ms.Sastry, AP Biology
Unit 3/Chapter 20
Summary of cloning gene of interest using plasmid vectors: goals 1 through 3
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Ms.Sastry, AP Biology
Unit 3/Chapter 20
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Goal 3 extension: Human genome project vs Craig Venter’s shotgun approach to sequence the entire
human genome. (Know how DNA is sequenced)
Goal 3 alternative – clone DNA, make huge numbers of copies of gene of interst without using a
plasmid/vector: PCR (polymerase Chain reaction) – very important.
Ms.Sastry, AP Biology
Unit 3/Chapter 20
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Goal 4: Identify and cut out gene of interest from the cloned libraries: DNA Gel electrophoresis:
Goal 4 extension: ‘RFLP’ – what is this and how is it used in gel electrophoresis (DNA fingerprinting) to
identify criminals/establish paternity/serve as marker for disease genes?
Ms.Sastry, AP Biology
Unit 3/Chapter 20
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Goal 5: make the human gene product
Gene chip/microarrays – what are these cool DNA tools?
Summarize in your own words how recombinant DNA technology can be used to make large amounts of human
growth hormone be produced in the milk of a cow:
Ms.Sastry, AP Biology
Unit 3/Chapter 20
Know the Vocabulary below: chapter 20
Gene of interest:
Restriction enzyme:
DNA Ligase:
Cloning DNA means:
Transformation:
Antibiotic selection:
Colony screening:
Probe:
Genomic library:
Human Genome Project:
PCR:
primer:
Gel electrophoresis:
Southern blotting:
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Plasmid:
Palindrome:
Sticky ends:
Vector:
Heat shock:
Ampicillin, Tetracyclin:
Marker/reporter:
cDNA:
DNA sequencing:
Shotgun approach:
Taq polymerase:
RFLP:
DNA Fingerprinting:
Transgenic organism: