Biological Mechanisms in Genetic Engineering and Protein Purification

Flashcards on key concepts and mechanisms related to genetic engineering and protein purification as discussed in the lecture.

Gene Isolation

The process of cutting out a desired gene using restriction enzymes.

Restriction Enzymes

Enzymes that recognize specific DNA sequences and cut them, creating ‘sticky ends’ for gene insertion.

Plasmid

A circular DNA molecule used to transport and replicate foreign genes within bacteria.

Ligation

The process of inserting a gene into a plasmid, where DNA ligase seals the sugar-phosphate backbone.

Transformation

The method of introducing a plasmid into bacteria to enable them to express foreign genes.

Heat Shock

A technique used to induce the uptake of DNA by bacteria through temperature changes.

Origin of Replication (ori)

A specific sequence where DNA replication machinery assembles to start the replication process.

Promoter

A DNA sequence that initiates transcription by binding RNA polymerase.

IPTG

A molecule used to activate gene expression by inactivating the lac repressor protein.

Ampicillin Resistance

A trait conferred by the presence of a beta-lactamase gene in plasmids, allowing bacteria to survive in ampicillin.

Affinity Chromatography

A technique that separates proteins based on their specific binding interactions with ligands attached to a stationary phase.

Size-Exclusion Chromatography

A method that separates molecules based on size by using porous beads in a chromatography column.

Ion-Exchange Chromatography

A separation method that relies on the charge characteristics of proteins to bind and elute them from a charged column.

Recombinant DNA

DNA that has been artificially made by combining DNA from different sources.

β-lactamase

An enzyme produced by resistant bacteria that breaks down ampicillin, rendering it inactive.

Selectable Marker

A gene included in plasmids that allows for the identification of successfully transformed cells.