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What is the process of recombinant protein expression?

Update time : 2020-10-22

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What is the process of recombinant protein expression?

Recombinant protein expression technology can be used to analyze gene regulation and study protein structure and function. Whether it is in vivo function research or large-scale production for structural research, as well as drug development in the field of biotherapy, protein expression technology is required.


The following are some knowledge points of protein expression summarized by General Biosystems.

General protein synthesis pathway:

DNA —>mRNA—> protein

The actual operation process is:

1. Clone or synthesize the target gene

2. Vector construction

3. Protein expression and purification

4. Mass expression and purification of protein (or none)


Clone or synthesize the target gene

A typical protein expression vector includes at least 4 main elements:


• Target gene expression box (including promoter and gene termination or poly (A) signal)

• The origin of replication of bacteria (ori)

• Antibiotic selection box for specific host (such as hygromycin B, mycophenolic acid, puromycin, blasticidin S, etc.)

• Antibiotic selection box for Escherichia coli (such as ampicillin, blasticidin S, carbenicillin, Zeocin selective antibiotic resistance, etc.)


Other components include:

• Multiple cloning site (MCS) (ie multi-restriction linker)

• Epitope tags

• Secretion signal

• Protease recognition site

• Internal ribosome insertion site (IRES)


Several common protein expression cases

Protein expression is the main content of many experimental subjects. Due to different research fields and various expression vectors, the characteristics of the protein that each person wants to express are extremely different. 

Especially some small partners are working on the subject for the first time. How to express the protein and how to arrange the experimental procedures is even more confusing.  

Here are the most commonly used recombinant protein expression schemes according to different expression vectors. 

Compare each protein expression case from the experimental steps, the time required, and the goals to be completed in each step. You can compare your own experimental content and hope it will help.

 

Commonly used protein expression (including cell line preparation):

• CHO stable cell line development

• HEK293 stable cell line development

• Prokaryotic protein expression

• Transient expression in mammalian cells

• Yeast protein expression

• Baculovirus and insect cell protein expression

Comparison of different protein expression

The completion time of the above experiment plan is the working process time of a more mature laboratory, and you can flexibly grasp it according to your own experiment arrangement. 

To accomplish the goal, you should also make appropriate additions and reductions based on your own subject requirements. 

If you have a relatively large-scale research topic, and protein expression is only part of the whole topic, General Biosystems can provide the above various protein expression services. 

General Biosystems' technical advantages:

Highly efficient expression vector

High density cell culture

Culture medium formula with independent property rights

>6,000 protein expression, purification and production experience

Inclusion body protein renaturation technology and soluble expression