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    生物資訊

    卓越性能,Mobitec巨大芽孢桿菌表達系統

    作者:admin 來源:本站 發布時間: 2019-04-16 19:06  瀏覽次數:
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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 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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