Protein Expression and Purification

These flashcards cover key concepts related to protein expression and purification from the lecture notes, helping students study for their exams.

What is a plasmid in genetic engineering?

A plasmid is a small, circular DNA molecule used to transfer genetic material into a host cell.

What is a fusion protein?

A fusion protein is a protein created by joining two or more genes that originally encoded separate proteins.

Why is affinity purification important?

Affinity purification allows for the isolation of a specific protein from a mixture by utilizing a tag that binds to a specific ligand.

What role does the fusion tag play in protein purification?

The fusion tag provides a means to easily isolate and purify the fusion protein from other cellular proteins.

What is IPTG and its significance in protein expression?

IPTG is an inducer used to initiate protein expression in systems that use inducible promoters.

What are potential disadvantages of constitutive protein expression?

Constitutive protein expression can lead to metabolic burden on the cell, slow growth, and protein toxicity.

What is chromatography?

Chromatography is a technique used to separate components of a mixture based on their interactions with stationary and mobile phases.

How does elution work in affinity chromatography?

Elution is the process of washing the bound protein off the beads by disrupting the interaction between the protein and the ligand.

What is the purpose of using spacer arms in affinity purification?

Spacer arms help enhance the binding affinity between the protein and the ligand by reducing steric hindrance.

Why is it important to purify proteins for vaccine use?

Purifying proteins prevents unwanted immune responses against other cellular proteins present in the mixture.