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生物资讯

卓越性能,Mobitec巨大芽孢杆菌表达系统

作者:admin 来源:本站 发布时间: 2019-04-16 19:06  浏览次数:
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性能参数

产品名称: 卓越性能,Mobitec巨大芽孢杆菌表达系统
英文名称: Bacillus megaterium Expression Systems
产品型号: kit
产品规格: kit
产品产地: 德国
品牌商标: MOBITEC
价    格: 面议元
 

产品详细描述

 

说明书(bmeg_handbook.pdf)

Features:

  • Stable, high-yield protein expression in Bacillus megaterium 
     
  • Ideal for both small and large-scale protein production
  • Tightly regulated and efficiently inducible xylose operon (up to 350-fold)
  • No alkaline proteases activity even up to 5 hours after induction
  • No endotoxins observed
  • Versatile cloning (extended polylinker, additional BsrG I site*)
  • Versatile production (intracellular or extracellular)
  • Versatile purification (native, 6xHis-tag, Strep-tag, Strep/6xHis double-tag)
  • Removeable tag versions (for TEV or Xa proteases) available New!
  • Compatible with all B. subtilis vectors

*BsrG I site is not included in all vectors.

The B. megaterium expression system provides a versatile and easy-to-handle tool for stable and high-yield protein production, both small and large scale.

B. megaterium has proven to be an excellent alternative host to E. coli for heterologous gene expression. Unlike other bacilli strains, proteolytic degradation by alkaline proteases is avoided. In addition, there are no endotoxins found in the cell wall.

High Protein Yield
As a result, protein yields are exceptionally high even if inexpensive substrates are used. For example, the proteins mutarotase (Mro) and glucose dehydrogenase (Gdh) have been accumulated up to 20% and 30% of the total soluble protein, respectively. Using the tightly regulated xylose operon, the genes were induced 130- to 350-fold without proteolysis.


Versatile System with a Wide Range of Vectors
MoBiTec provides a wide range of useful vectors for the B. megaterium system that are adaptable for most applications and protein purification methods. These include vectors carrying a secretion signal peptide sequence, a 6xHis-tag, a Strep-tag, and a Strep/6xHis double-tag. Additionally, all vectors contain the tightly regulated and highly inducible xylose operon promoter.

B. megaterium protoplasts specifically optimized for transformation 
The protoplasts supplied by MoBiTec are from B. megaterium strain WH320 developed by Prof. Dr. W. Hillen at the Institute of Microbiology in Erlangen, Germany. These protoplasts are prepared according to an optimized protocol resulting in the highest transformation efficiencies. 

For secretion vectors we offer protoplasts of strain MS941. Both strains are mutants of DSM319, where WH320 is a chemically mutant, while MS941 has a defined deletion in the gene of the major extracellular protease NprM.

Examples:

Selected Proteins successfully over-produced in Bacilli strains with our B. megaterium vectors:

  • β-Galactosidase (LacZ)1
  • Catabolite control protein (CcpA)2,3
  • Clostridium difficile toxin A4
  • Cobaltochelatase (CbiX)5
  • Dextransucrase6
  • Endolevanase (LevB)7
  • Glucose dehydrogenase (GdhA)1
  • Heat shock protein (HPr) from PTS (phosphotransferase sugar transport system)8
  • Human single-chain urokinase-like plasminogen activator (rscuPA)1
  • Levansucrase9
  • Mutarotase (Mro)1
  • Neopullulanase10
  • Translocation ATPase of the preprotein translocase (SecA)11
  • Trehalose repressor (TreR)12

 

 

order#
description
amount
BMEG02
Bacillus megaterium protoplasts ready for transformation (strain WH320;1)
Material is sufficient for 4 transformations plus control experiment.
5 x 500 µl
BMEG50
Bacillus megaterium protoplasts, strain MS941
5 x 500 µl
BMEG31
pC-His1623hp shuttle vector, C-term. 6xHis-tag; lyophilized DNA
10 μg
BMEG32
pN-His-TEV1623hp shuttle vector, N-term. His-tag,TEV-site
10 μg
BMEG30
p3STOP1623hp, shuttle vector, lyophilized DNA
10 μg
BMEG33
pSPLipA-hp, shuttle vector with signal sequence, lypophilized DNA
10 μg
BMEG34
pSPYocH-hp, shuttle vector with signal sequence, lypophilized DNA
10 µg
BMEG03
pWH1520 shuttle vector, original; lyophilized DNA
5 µg
BMEG10
pMM1522 shuttle vector, improved; lyophilized DNA
10 µg
BMEG11
pMM1525 shuttle vector with signal sequence; lyophilized DNA
10 µg
BMEG12
pHIS1522 shuttle vector, 6xHis-tagged; lyophilized DNA precursor of BMEG20
10 µg
BMEG13
pHIS1525 shuttle vector with signal sequence; 6xHis-tagged; lyophilized DNA
10 µg
BMEG14
pSTREP1525 shuttle vector with signal sequence; Strep-tagged; lyophilized DNA
10 µg
BMEG15
pSTREPHIS1525 shuttle vector with signal sequence;
Strep/6xHis double-tagged; lyophilized DNA
10 µg
BMEG20
pC-His1622 shuttle vector, C-term. 6xHis-tag; lyophilized DNA
10 µg
BMEG21
pC-Strep1622 shuttle vector, C-term. Strep-tag; lyophilized DNA
10 µg
BMEG22
pN-His-TEV1622 shuttle vector, N-term. His-tag,TEV-site
10 µg
BMEG23
pN-Strep-TEV1622 shuttle vector, N-term. Strep-tag,TEV-site
10 µg
BMEG24
pN-StrepXa1622 shuttle vector, N-term. Strep-tag; Xa site; lyophilized DNA
10 µg
BMEG25
pSTOP1622, shuttle vector, lyophilized DNA
10 µg
BMEGT702
Bacillus megaterium protoplasts, strain MS941, pretransformed with pT7-RNAP
5 x 500µL
BMEGT701
Bacillus megaterium high yield T7 gene expression kit, 
includes pretransformed protoplasts BMEGT702 (5 x 500µl), pPT7 cloning vector and pPT7-GFP control vector (vectors lyophilized, 10 µg each)
Kit
BMEGT710
Bacillus megaterium pPT7 cloning vector, lyophilized
10 µg
shipped at RT, protoplasts shipped on dry ice 
store lyophilized vectors at 4°C, reconstituted vectors at -20°C, protoplasts at -70°C
All vectors are E. coli / B. megaterium shuttle vectors
Please note that protoplasts of Bacillus megaterium have a limited shelf life of some months only, depending on the date of production. Therefore purchase and use of protoplasts must be thoroughly planned.

 

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网址:www.bitebo.com

 

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