A Leap Forward in Protein Production Visit our Website Today! www.takara-bio.eu Brevibacillus Expression System Protein Research Products High Yields of Disulfide Bonded and Secreted Proteins High Efficiency Secretion of Large Quantities of Proteins Proteins are Produced in Biologically Active Forms Good at Intracellular Production as well
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A Leap Forward in Protein Production
Visit our Website Today!www.takara-bio.eu
Brevibacillus Expression
System
Protein Research Products
High Yields of Disulfide Bonded and Secreted Proteins
High Efficiency Secretion of Large Quantities of Proteins
Proteins are Produced in Biologically Active Forms
α-amylase B. licheniformis 3.7Sphingomyelinase B. cereus 3Xylanase B. halodurans 0.2CGTase B. macerans 1.5Chitosanase B. circulans 1.4Hyper thermo-stable protease A. pernix 0.1Hyper thermo-stable nuclease P. horikoshii 0.7PDI human 1AntigensSurface antigen E. rhusiopathiae 0.9Surface antigen T. pallidum 0.8CytokinesEGF human 1.5IL-2 human 0.6NGF mouse 0.2IFN- γ chicken 0.5TNF-α bovine 0.4GM-CSF bovine 0.2GH flounder 0.2
Proteins Origins Production( g/L )Enzymesα-amylase B. licheniformis 3.7Sphingomyelinase B. cereus 3.0Chitosanase B. halodurans 0.2CGTase B. macerans 1.5Chitosanase B. circulans 1.4Hyperthermostable protease A. pernix 0.1Hyperthermostable nuclease P. horikoshii 0.7PDI human 1.0AntigensSurface antigen E. rhusiopathiae 0.9Surface antigen T. pallidum 0.8CytokinesEGF human 1.5IL-2 human 0.6NGF mouse 0.2IFN- γ chicken 0.5TNF-α bovine 0.4GM-CSF bovine 0.2GH flounder 0.2
Recovery of Dehydrogenase activity is 10 to 20 times higher when expression is performed using the Brevibacillus expression system versus expression using E.coli BL21 (DE3). In conclusion, the Brevibacillus expression system produces a higher yield of active protein.
16h induction
- induction 2SLNm TMNm
SPP
PP Gene
Expressionvector
Expressionvector
+Ligase
Stopcodon
SPcleavagesite
Gene
PCRFragment
:RestrictionSite
:Promoter
:Secretionsignalpeptide
:SDsequence
SP
SP
TaKaRa Bio recommends
High fidelity polymerase for PCR amplification of the target gene:PrimeSTAR HS DNA polymerase (R010A; R044A or R040A)
For efficient synthesis of long cDNA fragments: PrimeScript RTase (2680A or 6110A)
For protein purification:Talon Resin (635501 or 635606) or His60 (635659)
For insert cDNA cloning :DNA Ligation Kit <Mighty Mix> (6023) or DNA Ligation Kit Ver.2.1 (6022) and Takara’s restriction enzymes
Product Name Catalog # Quantity Brevibacillus Expression system tik 001 BHKit Components
Expression vectors pNY326 DNA gµ 01111 BHpNCMO2 DNA gµ 01211 BHControl Vector pNCMO2-BLA DNA gµ 1311 BHCompetent cells (for electroporation)Brevibacillus choshinensis Electro-cells HB 115 100 µl x 10 tubes
Brevibacillus Expression System Kit
License Notices [L16] His-Tag Sequence This product is covered by the claims of U.S. Patent No. 5,284,933, 5,310,663 and their foreign counterpart patent claims. Protein Purification Technology of His-Tag used in some of pCold vectors and Brevibacillus expression vectors is licensed from Hoffmann-La Roche, Inc., Nutley, NJ and/or Hoffmann-La Roche Ltd., Basel, Switzerland and is provided only for the use in research. Information about licenses for commercial use is available from QIAGEN GmbH, Qiagen Strasse 1, D-40724 Hilden, Germany. [L39] Brevibacillus Expression System 1. B. choshinensis SP3, pNY326, pNCMO2, pNCMO2-BLA, pNC-HisT, pNC-HisF, pNC-HisE, pNI and pNI-His were developed and manufactured by Higeta Shoyu Co., Ltd. and sold by TAKARA BIO INC. 2. The use of this product is allowed for academic research only. You have to conclude a license agreement if you use this product or the products produced by using this product for commercial purpose. Contact us if you would like to ask any question about license etc. If any questions are received, either Higeta Shoyu Co., Ltd. or an agent for it will answer. TaKaRa Bio Inc. Tel +81-77-543-7247 Fax +81-77-543-9254 3. All rights of intellectual property related to this product are owned by Higeta Shoyu Co., Ltd. This product is covered by the following Japanese patents and corresponding international patent : Japanese patents, 2103774, 3696322, 3734593, 3753945, PCT/JP2004/016912 (international application based on PCT treaty), 3433807. 4. This product, the components or the derivatives of this product, or the products produced by using these items can not be transferred (distributed freely or sold) to third party. (When you purchase this product, you submit the license agreement to us.)Notice: All products are intended to be used for research purpose only. They are not to be used for drug or diagnostic purposes, nor are they intended for human use. They shall not to be used products as food, cosmetics, or utensils, etc. Takara products may not be resold or transferred, modified for resale or transfer, or used to manufacture commercial products without written approval from TAKARA BIO INC.
All shuttle vectors between B. choshinensis andE.coli contain the P2 promoter, which is one of the five promoters that control transcription of the cell wall protein gene (HWP). This promoter does not work in E.coli but in B. choshinensis. The P2 promoter is quite robust and efficiently produces proteins.
The pNY326 vector* is maintained more stable than pNC or pNI vectors in the host cells due to much weaker promoter activity and to its smaller size (3.4 kb). Sometimes it works better than shuttle vectors, and is useful for a large scale production. The pNY326 vector must be constructed by the one step method using B. choshinensis. Brevibacillus choshinensis Electro-Cells(105 transformants/ug DNA) is used as the transforma-tion host.