General aspects of

heterologous protein
expression
General aspects of heterologous protein
expression

• Heterologous expression is the introduction of either complementary

DNA (cDNA) or RNA (cRNA) encoding for a protein of interest from

one species into the cell of another species, such that the hosts'

cellular machinery expresses the foreign protein.

• Plants are a promising expression system for the production of heterologous proteins,

especially therapeutic proteins.

• Currently the majority of therapeutic proteins are produced in mammalian cell lines

or bacteria.

• In a few cases insects, yeast and fungi have been developed for production of human

proteins.
• Heterologous expression refers to the expression of a gene or part of

a gene in a host organism that does not naturally have the gene or

gene fragment in question.

• Insertion of the gene in the heterologous host is performed by

recombinant DNA technology.

• An efficient expression vector requires a strong promoter with a high

affinity for the RNA polymerase to increase the transcription of the

heterologous gene.

• A strong and adjustable promoter can control the transcription of the

target gene in a certain period.

• Heterologous protein expression is an important method for analyzing

cellular functions of proteins, in genetic circuit engineering and in

overexpressing proteins for biopharmaceutical applications and

structural biology research

Examples
Products Strains

Insulin-like growth factor-1

E. coli W3110
fusion protein

Insulin-like growth factor-2

E. coli BL21(DE3)
(IGF-2)

Human granulocyte colony-

E. coli BL21(DE3)
stimulating factor (hGCSF)

Growth hormone E. coli W3110

Advantages

• Heterologous expression may provide higher enzyme yields and may

permit the production of laccases with desired properties, such as

different substrate specificities, or improved stabilities for industrial

applications
• Traditional strategies for recombinant protein expression

involve transfecting cells with a DNA vector that contains the

template and then culturing the cells so that they transcribe and

translate the desired protein. Typically, the cells are then lysed to

extract the expressed protein for subsequent purification.

Methods of protein expression

• Several methods are used, such as sonication, repeated freezing,

thawing, homogenization through high pressure or grinding, or

permeabilization through detergents or enzymes. Further, this

method releases proteases during the cell lysis, enabling the onset of

digestion of proteins in the solution

Heterologous and Homologous protein
expression

• Heterologous expression is when you express a gene in either a

different specie, or cell type, than it originates. Homologous would

then be the over-expression of a gene in a system from where it

originates.
Gene expression steps
• The gene expression steps are:
• Transcription.
• mRNA processing.
• mRNA export.
• Translation.
• Protein folding.
• Protein translocation.
Factors affecting gene expression

• Various factors, including genetic makeup, exposure to harmful

substances, other environmental influences, and age, can affect

expressivity. Both penetrance and expressivity can vary: People with

the gene may or may not have the trait and, in people with the trait,

how the trait is expressed can vary.