Lab Final Flashcards
What is Microplusin?
Antimicrobial Peptide
What are antimicrobial peptides?
Peptides that help attack bacteria
Is the antimicrobial peptide that we are working with anionic or cationic?
Catatonic
What organism is Microplusin derived from?
Isolated from the cell-free hemolymph and egg of the tick Boophilus microplus
What is PRSETA?
It is a vector used for high level protein expression
PRESTA has an antibiotic resistance gene to what antibiotic?
Ampicillin
What are competent cells?
Cells that can uptake foreign DNA
Define transformation.
Foreign DNA introduced into a cell
What type of E. coli were used the transformation?
BL21
What media was used to grow the transformed cells?
LB Media
What antibiotic was used?
Ampicillin
Describe the Lac Operon
Cluster of genes used to regulate lactose
What is heat shock and why is it performed?
To form pores in the cell for DNA uptake
What is IPTG? How does it work?
IPTG is an inducer that mimics lactose and is used to induce protein expression
For how many hours did we incubate the induced protein after induction with IPTG?
4 hours
True or False. In order to express a gene that is controlled by the T7 promoter, you need to add IPTG to block the function of the lac repressor.
True
Name the method for colorimetric detection and total quantification of protein.
BCA assay
How does the BCA assay work? What does BCA stand for?
bicinchoninic acid assay; works by measuring the level of amino acid composition of the protein.
BCA is what type of reaction? Explain.
Colometric measure the absorption
True or False. The total quantification of the protein was measured in absorbance
True
What does SDS-PAGE stand for? How does it work?
Polyacrylamide gel electro
What is SDS? What does it do to proteins?
Sodium deodecyl Sulfate gives a negative charge to proteins
What are the metals used for greatest affinity for histidine?
Nickel
How many histidine does PRSETA have?
6
