Week 8 Review Flashcards
What are some examples of biotechnology?
Bread, beer/wine, cheese, yogurt
Antibiotics
Pharmaceuticals
Genetically modified organisms (GMOs)
What are the requirements (e.g. plasmid), and steps to produce a recombinant piece of DNA?
A restrict enzyme cuts out a gene of interest
Ligase combines human DNA with the gene of interest
When the bacteria divide, the recombinant bacteria gain the ability of the gene of interest
How are restrictions involved in recombinant DNA? What do they produce?
Restriction enzymes cut specific parts of a DNA sequence, producing a free gene of interest.
The gene of interest will combine with a plasmid via ligase and produce a bacterium that gains the ability of the gene of interest.
How is the recombinant DNA introduced in a cell and how is it selected?
Recombinant DNA is introduced in a cell by plasmids that act as a vector for foreign DNA
Plasmids contain antibiotic-resistant genes so the bacteria are grown on a specific antibiotic, and if they’re resistant, that means they have the plasmid
What are the steps in cloning a gene?
DNA is extracted from the organism and is cut into fragments and inserted into plasmids
The bacteria are transformed with plasmids (vectors that carry foreign DNA)
The bacteria replicate, producing colonies of clones
What was the first commercial genetically engineered product?
Insulin (Genentech 1982)
- Insulin gene cloned into plasmid and expressed in E.coli
- Large cultures were grown and the protein was extracted
What are the two forms of DNA profiling? What is the purpose and what are the steps involved in the two forms?
Restriction Fragment Length Polymorphism (RFLP): allows to track infectious diseases
- DNA cut with restriction enzymes
- Different individuals produce different fragments
- Separates based on length
- Use a probe to digest and separate a specific region of DNA on gel electrophoresis to observe mutated DNA and compare to normal DNA
Polymerase Chain Reaction (PCR): small samples of DNA are amplified
- Denature DNA (heat up)
- Anneal primer (stick)
- Extension (add DNA pol to copy DNA)
- Cycle repeated
What are other uses for PCR?
Detect the presence of infectious agents
- qPCR provides rapid detection of M. tuberculosis and takes 6 weeks to culture
What does CRISPR stand for? How is used by bacteria and how has it been applied to gene editing?
CRISPR-CAS 9: used naturally in prokaryotes
- Allows bacteria to “remember” viruses
Used in biotechnology to edit genes of interest
Review CRISPR/Cas
CRISPR-Cas 9 Immunity:
90% archaea
33% bacteria
How it works:
- The Cas9 protein forms a complex with guide RNA in the cell
- Complex attaches to matching genomic DNA sequence adjacent to a spacer
- The cas9 RNA complex cuts double strands of DNA
- Programmed DNA may be inserted at the cut
