Bacillus Megaterium Classic Protein Expression System

The Classic xylose operon-regulated Bacillus megaterium expression service is now available at Creative BioMart. In order to provide high-quality service, we offer various plasmids and strains for customers to choose from.

Service Principle

We use shuttle vectors of Escherichia coli and Bacillus megaterium. After the GOI is inserted, the vector is replicated in E. coli and transformed into Bacillus megaterium protoplasts. (We usually use strain DH10B. If you have any special needs, please let us know in advance.)

After transformation, we use xylose to induce the expression of the recombinant gene. We will collect a small sample of culture samples before and after induction to determine the successful overexpression of the target gene.

Here is Creative BioMart's classic Bacillus megaterium protein expression procedure and deliverable materials.

1. Selection of expression system (if the customer provides the expression strain, this step will be omitted. Only the optional host strain of expression plasmid will be provided.)
Host strains and shuttle vectors selection	1. Classic secretory expression system 2. Classic non-secretory expression system
2. Shuttle vector construction	Deliverable
Gene optimization Subcloning design Synthetic gene Cloning strategies selection Subclone the GOI to shuttle vector	1. Gene synthesis report 2. Sequencing validation report
3. Obtain the recombinant shuttle vector
Recombinant shuttle vector cultured in E.coli strain Shuttle vector extraction	1. Cloning strain containing expression plasmid (optional) 2. shuttle plasmid 3. Plasmid instructions
4. Analysis and identification of protein expression test
Transform the shuttle vector into Bacillus megaterium protoplasts Expression test Expression analysis and identification Expand scale	1. Positive expression strains 2. Small test expression process report 3. Images and spectrums obtained from the analysis 4. Small test expression summary
5. Protein expression and purification
Protein affinity purification (extracellular or intracellular) Identify purified proteins using WB	1. Protein purification process report 2. Original picture of protein purification 3. Delivery of protein 4. Western blot (labeled-antibody) 5. Protein fingerprint (optional) 6. Molecular weight of protein mass spectrometry (optional)
6. Scale-up culture in fermentor
Amplification Culture (10-20l) Amplification Culture (above 50L)	1. Protein purity identification report 2. Finished protein (agreed price according to the results of expression test)

Vectors and Strains

Here are some shuttle vectors and strains that available at Creative BioMart.

Vector	Secretion	N-terminal tag		C-terminal tag		Tag cleavable
		6 x His	Strepll	6 x His	Strepll
pWH1520	-	-	-	-	-	N/A
pWH1522	-	-	-	-	-	N/A
pWH1525	Yes	-	-	-	-	N/A
pHIS1522	-	-	-	Yes	-	-
pHIS1525	Yes	-	-	Yes	-	-
pSTREP1525	Yes	-	Yes		-	Yes
pSTREPHIS1525	Yes	-	Yes	Yes	-	Yes (Strepll)
pSTOP1622	-	-	-	-	-	N/A
pC-His1622	-	-	-	Yes	-	-
pC-Strep1622	-	-	-	-	Yes	-
pN-His-TEV1622	-	Yes	-	-	-	Yes
pN-Strep-TEV1622	-	-	Yes	-	-	Yes
pN-Strep-Xa1622	-	-	Yes	-	-	Yes

Creative BioMart offers three different B. megaterium strains: WH320, YYBm1 and MS941.

Service Features

• Customized expression scheme, according to the nature of the product, using different strategies
• A variety of optional vectors and strains for customers to choose from
• An experienced team that has successfully completed protein expression for hundreds of customers
• One-stop services

Related Service

Didn't find what you need? You may be interested in the following services.

• Bacillus megaterium High Yield Expression system
• Bacillus megaterium T7-RNA Polymerase Expression system

Customers who need our assistance are more than welcome to contact us or submit an online inquiry.