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性能參數

產品名稱：卓越性能，Mobitec巨大芽孢桿菌表達系統

英文名稱： Bacillus megaterium Expression Systems

產品型號： kit

產品規格： kit

產品產地：德國

品牌商標： MOBITEC

價格：面議元

產品詳細描述

說明書(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
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	pSP_LipA-hp, shuttle vector with signal sequence, lypophilized DNA	10 μg
BMEG34	pSP_YocH-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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