biotech / dna Flashcards
What is biotechnology?
The use of living organisms in order to produce a desired product or effect.
Biotechnology encompasses various processes, including fermentation and genetic manipulation.
List some reasons for manipulating DNA.
- Forensic Science
- Agriculture and Crop research
- Human Health
- Gene therapy
- Designer drug therapies
- Bioremediation
- Bioinformatics
These applications highlight the importance of DNA manipulation in various fields.
What is recombinant DNA?
DNA that is created by combining the DNA of at least two different sources into one DNA molecule.
Recombinant DNA technology is a fundamental technique in biotechnology.
What are restriction endonucleases?
Enzymes that cut DNA at specific sequences.
They are crucial for creating recombinant DNA.
What is gel electrophoresis used for?
To separate DNA fragments based on their size.
This technique helps visualize DNA after it has been manipulated.
What are plasmids?
Small circular DNA molecules used as vectors in genetic engineering.
Plasmids are often used to transfer genes into host cells.
What are target genes?
Specific genes that are intended to be manipulated or expressed in a genetic engineering process.
Identifying target genes is essential for successful DNA manipulation.
What are fusion proteins?
Proteins created by joining two or more genes that originally coded for separate proteins.
Fusion proteins can have novel properties and functions.
True or False: The concept of manipulating DNA is a recent development.
False.
DNA manipulation has existed for thousands of years, starting with simple interbreeding of species.
What type of restriction enzymes are most commonly used for DNA analysis and genetic engineering?
Type II restriction enzymes
They have unique nucleotide sequences at which they cut DNA.
What kind of sequence is a recognition sequence?
palindromic sequence (with 6 base pairs)
Some may recognize four or eight base pair sequences.
What type of DNA ends can result from different cutting patterns of restriction enzymes?
sticky ends
sticky ends have short single-stranded overhangs.
How do sticky ends facilitate DNA manipulation?
Through complementary base-pairing
This allows the ends to potentially join together again.
What do restriction enzymes generate to characterize a specific dna molecule?
DNA fingerprint
They create discrete fragments resolved by gel electrophoresis.
What is the result of enzymes that cut in the middle of the recognition sequence?
Blunt ends
This results in clean cuts without overhangs.
What technique is used to separate a mixture of molecules through a gel in an electric field?
Gel electrophoresis
This technique separates DNA fragments based on size.
what causes the DNA fragments to move through the gel?
DNA is negatively charged while the other end of the chamber is positive so it attracts to the positive end
what do restriction enzymes defend against?
bacteriophages
What is the size range of plasmids?
1000 to 200,000 base pairs
The size of plasmids can vary significantly.
What is a selection site in the context of plasmids?
A gene for antibiotic resistance that allows bacterial cells to thrive
Common antibiotics include tetracycline and ampicillin.
What type of relationship exists between bacteria and plasmids?
Endosymbiotic relationship
Both bacteria and plasmids benefit from this relationship.
What is a restriction map?
A preparation that indicates where specific enzymes will cut the plasmid
This is made possible due to the well-known DNA sequence of plasmids.
What is the cloning region in plasmids?
The area high in restriction sites where genes of interest are inserted
This region is crucial for cloning procedures.
What is the first step in cloning DNA using plasmids?
Cut DNA to be cloned with the same restriction enzyme used on the plasmid
This ensures compatibility for ligation.
True or False: Plasmids can be replicated and their proteins expressed using enzymes and ribosomes found in the bacterial cell.
True
This capability is one of the reasons plasmids are valuable in genetic engineering.
What are the steps involved in creating a recombinant plasmid?
- Cut plasmid with restriction enzyme
- Cut DNA to be cloned with the same enzyme
- Ligate the two DNAs to form a recombinant molecule
- Introduce into host cell
- Select for cells carrying recombinant plasmids
Selection can be done using antibiotics or color indicators.
