Genetics 133 Exam 3 UW-
Madison with 100% correct
answers 2025
GMOs are - answer plants with one or more genetic characteristics
inserted into genome using biotechnology
GMOs are NOT - answer plants improved through artificial selection
processes like cross breeding, hybridization, or mutagenesis
Creating the Roundup Ready Soybean - answer
Thegene,CP4EPSPS,wasfoundinamicrobe (bacteria)
The gene was isolated and extracted from the microbe
The gene was inserted into aplasmid
The gene was inserted into the soybean genome using particle gun
bombardment and plant transformation
The plant with the new gene was grown and observed to determine if the
insertion was successful
Field and safety test were done before it was approved for commercial use
Recombinant DNA technology - answer the joining together of DNA
molecules from two different species to produce new genetic combinations
that are of value to science, medicine, agriculture and industry
Two ways to isolate and amplify your gene of interest - answer in vivo
and in vitro
In vivo - answer in the living
• It indicates that an experiment is being done in conditions that precisely
mirror those existing innature,usuallywithinanatural organism
,In vitro - answer in a test tube
• Highly controlled environment, much easier to set up
• May not accurately predict the conditions inside aliving organism
Overview of isolation and amplification of a gene of interest in vivo -
answer 1) IsolatingDNA-Beginwith sampleof DNA containing geneof
interest (donor DNA)
2) CuttingDNA-Donor DNA cut upwith restriction
enzymes(molecularscissors)
3) JoiningDNA-Inserted into vector chromosometo makerecombinantDNA
4)Amplifying Recombinant DNA-Transferred into bacterial cells to be
amplified by bacterial cell replication
5)DNAcloning-singlecellcontaining recombinant
DNAbecomescloneofidentical cells
Genomic DNA can be cut up before cloning using - answer restriction
enzymes, creates sticky ends or blunt ends
To cut genomic DNA into smaller fragments suitable for cloning use -
answer bacterial derived restriction enzymes
Restriction enzymes cut at - answer specific DNA sequences, restriction
sites
Same restriction enzyme will cut - answer DNA same every time
Each enzyme cuts - answer a different DNA palindromic sequence
Overview of isolation and amplification of a gene of interest in vitro -
answer • PCR used to amplify gene or DNA of interest (target DNA)
• Primers specific to sequence of target DNA usedto guide replication
process by DN A polymerase
• Repeatedcyclesresultinlarge quantities of target DNA
, Four components are required for PCR - answer PCRrequires four
components to successfully clone gene of interest:
1) DNA template-make copies from
2) Primers-used to prime DNA polymerase
3) dNTPs- deoxynucleotide triphosphates (adenine, guanine, cytosine and
thymine bases)
4) Heat-tolerant polymerase-to perform DNA replication
The polymerase chain reaction amplifies selected regions of DNA in vitro -
answer 1) Mix four components together in a tube
2) Heat denature (separate) the DNA double strands at 95°C
3) Cool to allow primers to anneal (temp depends on GC content)
4) Heat to allow DNA synthesis with heat-tolerant DNA polymerase
5) Synthesis occurs from primers
Three steps involved in PCR are repeated again for - answer 25-35 cycles
to amplify your DNA a billion-fold,
Double-stranded DNA molecules identical to parent are produced
One cycle-three steps-denaturation, annealing, synthesis
Repeat cycles results in exponential growth
Each cycle ~5 minutes
Amplification to a billion-fold within 2.5 hours
How do we use amplified DNA? - answer • Gene isolation for basic
biological research
• Gene therapy to treat human diseases
• Productionofherbicidesandpesticidesbycropplants
• Example:Insulinfor diabetes
• Body does not produce enough insulin (Type I diabetes) to control blood
sugar levels
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