Protein Induction Theory and Recombinant Enzyme Production

These flashcards cover key terms and concepts related to the Protein Induction Theory and the production of recombinant human enzymes in E. coli, emphasizing the mechanisms and components involved in the process.

Dihydrofolate reductase (DHFR)

A recombinant human enzyme produced in E. coli used for various studies.

Glutathione S-transferase (GST) tag

A fusion protein tag that helps maintain the solubility and activity of proteins.

Polyhistidine (His) tag

A fusion protein tag that allows for purification using nickel columns.

BL21(DE3) E. coli

A genetically modified bacterial strain containing the T7 RNA polymerase gene.

T7 RNA polymerase

An enzyme that recognizes the T7 promoter and transcribes genes on a plasmid.

Lac operon

A system that controls the transcription of the T7 RNA polymerase and GST-DHFR-His genes.

Lac repressor

A protein that binds to the promoter and inhibits transcription before induction.

IPTG

A small molecule that mimics lactose and induces protein expression by binding the lac repressor.

Inducible system

A system that allows for controlled expression of a gene in response to an external signal.

Fusion protein

A protein created by joining two or more genes that originally coded for separate proteins.

SDS-PAGE

A method used to separate proteins based on their molecular weight during analysis.

OD600

Optical density measurement at 600 nm used to estimate bacterial growth in culture.

Nickel column purification

A technique that utilizes His tags to purify proteins by binding them to nickel ions.

Protein induction

The process of initiating the expression of a protein from a gene by removing transcription inhibition.