Lecture

Protein Expression Systems

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In course

CH-319: Experimental biochemistry and biophysics

A 7-week long (4+8 h) experiment where you plan and construct a fluorescent sensor protein starting from DNA bricks. The protein will be expressed in and purified from E.coli, characterized by biochemical and spectroscopic methods. Your report will be in ...

Description

This lecture covers the process of protein expression, from vector design to the selection of expression systems. It discusses the use of E. coli BL21 (DE3) for protein expression, vector design considerations, and methods to increase protein solubility. The lecture also explores the advantages and disadvantages of using E. coli as an expression system, as well as other common systems like mammalian, insect, yeast, bacterial, and cell-free systems. Different fusion partners, selection markers, and expression vectors are presented, along with practical examples of protein expression in E. coli and insect cells.

Instructor

Rudolf Hovius

Expertise Â•Protein structure, expression & purificationÂ•Molecular interaction & receptor activationÂ•Fluorescence imaging & spectroscopySignalling & sensing Mission => Teaching: Lecture courses, tutorials and practicals in biochemistry, biophysical chemistry of membranes and signalling processes. Participating to the Commission d'Enseignement and Commission de l'Ecole Doctorale of Chemistry and Chemical Engineering.=> Research interests: Protein function and structure, Membrane protein (especially recepto

Official source

Related concepts (32)

Expression vector

An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for gene expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene. Expression vectors are the basic tools in biotechnology for the production of proteins. The vector is engineered to contain regulatory sequences that act as enhancer and promoter regions and lead to efficient transcription of the gene carried on the expression vector.

Gene expression

Gene expression is the process by which information from a gene is used in the synthesis of a functional gene product that enables it to produce end products, proteins or non-coding RNA, and ultimately affect a phenotype. These products are often proteins, but in non-protein-coding genes such as transfer RNA (tRNA) and small nuclear RNA (snRNA), the product is a functional non-coding RNA.

Protein production

Protein production is the biotechnological process of generating a specific protein. It is typically achieved by the manipulation of gene expression in an organism such that it expresses large amounts of a recombinant gene. This includes the transcription of the recombinant DNA to messenger RNA (mRNA), the translation of mRNA into polypeptide chains, which are ultimately folded into functional proteins and may be targeted to specific subcellular or extracellular locations.

Regulation of gene expression

Regulation of gene expression, or gene regulation, includes a wide range of mechanisms that are used by cells to increase or decrease the production of specific gene products (protein or RNA). Sophisticated programs of gene expression are widely observed in biology, for example to trigger developmental pathways, respond to environmental stimuli, or adapt to new food sources. Virtually any step of gene expression can be modulated, from transcriptional initiation, to RNA processing, and to the post-translational modification of a protein.

Yeast artificial chromosome

Yeast artificial chromosomes (YACs) are genetically engineered chromosomes derived from the DNA of the yeast, Saccharomyces cerevisiae, which is then ligated into a bacterial plasmid. By inserting large fragments of DNA, from 100–1000 kb, the inserted sequences can be cloned and physically mapped using a process called chromosome walking. This is the process that was initially used for the Human Genome Project, however due to stability issues, YACs were abandoned for the use of Bacterial artificial chromosome The bakers' yeast S.

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