bioluminescence test Flashcards
What are proteins, and what roles do they play in cellular functions?
Proteins are complex molecules made up of amino acids. They help with the structure, function, and regulation of cells.
Describe the structure of a protein and the levels of protein organization.
Primary - sequence of amino acids
Secondary - Local spatial arrangement of a polypeptide’s backbone
Tertiary - diff parts ofd protein chain begin to fold into a shape
Quaternary - the arrangement of four protein subunits coming together to form a functional complex
What is an enzyme, and how does it function as a catalyst in biochemical reactions?
Enzyme - catalyst of chemical reactions
Reduces activation energy needed and speeds up the reaction
Explain the significance of enzyme-substrate specificity in biochemical reactions.
Ensures correct process is being completed
Determines a products efficiency preventing side effects
Speeds up reactions
Explain the concept of the active site of an enzyme and its role in catalysis.
An active site is a pocket on an enzyme where the substrates bind, where the reaction takes place.
What is the difference between competitive and non-competitive inhibition of
enzyme activity?
Competitive inhibition occurs when a molecule competes with the substrate to bind to the active site of the enzyme.
Non-competitive inhibition occurs when an inhibitor binds to the enzyme at a location other than the active site and alters the structure of the enzyme.
Describe the process of cloning a gene into a plasmid vector. What are the key steps and enzymes involved?
- Allow bacteria to transform n take in the gene (need DNA ligase)
- Let bacteria grow n reproduce
- lyse the bacteria
- Extract plasmid
What is the role of restriction enzymes in the cloning process?
It cuts one gene from a sequence of genes (DNA)
Explain the principle of PCR and its importance in molecular biology.
Pcr is used to copy specific segments of DNA strands. Gene analysis/isolation
Describe the three main steps of a PCR cycle and the purpose of each step.
Denaturation: Separates the DNA into single strands.
Annealing: Allows primers to bind to complementary sequences flanking the target region.
Extension: DNA polymerase synthesizes new DNA strands using the primers as templates.
What are the essential components required for a PCR reaction?
Dna template
Primer
Dna polymerase
nucleotides
buffer
What is Gibson Assembly, and how does it differ from traditional cloning methods?
a way of cloning and connecting many DNA fragments together in a single step. It differs because you assemble a fragment of DNA together to create one strand.
Describe the main steps involved in Gibson Assembly.
Design Overlaps: Make sure each piece of DNA has matching ends that overlap with the ends of the other pieces.
Mix DNA Pieces: Put all the DNA pieces together in a tube.
Add Enzymes: Add special proteins (enzymes) that will help join the overlapping ends.
Incubate: Let the mixture sit for a while so the enzymes can work.
Joined DNA: The result is a single piece of DNA made from all the original pieces.
Why is E. coli commonly used in genetic engineering and biotechnology research?
First species of bacteria to be cultivated in lab ; Can be propagated extremely easy and grows exponentially quickly
Discuss the role of antibiotic selection in identifying successfully transformed E. coli cells.
because of the rapid reproduction.
