pN-Strep-Xa1622 BioVector NTCC质粒载体菌种细胞基因保藏中心

pN-Strep-Xa1622

Vectors of the 1520 series pWH1520 Original shuttle vector pWH1520 (Rygus and Hillen, 1991). Features as described above. pMM1522 Like pWH1520 with additional restriction site BsrGI upstream of the ATG. The BsrGI site can be used to clone the gene of interest with its native start codon (corresponding to the native N-terminus of the protein of interest). The vector contains a stop codon downstream of the ATG (> 120 bp). pMM1525 Like pMM1522 with additional signal sequence for protein secretion (SPlipA) upstream of the MCS (the native lipA encodes for an extracellular esterase). pHIS1522 Like pMM1522 with additional sequence for C-terminal 6xHis-tag fusion (including stop codon). pHIS1525 Like pMM1525 with additional sequence for C-terminal 6xHis-tag fusion (including stop codon). pSTREP1525 Like pMM1525 with additional sequence for N-terminal Strep-tag fusion (cleavable with Factor Xa protease) and stop codon downstream of the MCS. Note: this vector does not contain BsrGI restriction site. pSTREPHIS1525 Like pMM1525 with additional sequences for N-terminal Strep-tag (cleavable with Factor Xa protease) and C-terminal His-tag fusion (including stop codon). Note: this vector does not contain a BsrGI restriction site. Vectors of the 1622 series pSTOP1622 Size-reduced pMM1522 variant with a stop codon directly downstream of the MCS. pC-His1622 Size-reduced pHIS1522 variant with sequence for C-terminal 6xHis-tag fusion (including stop codon right downstream of the tag). pC-Strep1622 Size-reduced pHIS1522 variant with sequence for C-terminal StrepII-tag fusion (including stop codon directly downstream of the tag). pN-His-TEV1622 Like pSTOP1622 with additional sequence for N-terminal 6xHis-tag fusion, cleavable with TEV (tobacco etch virus) protease.
pN-Strep-TEV1622 Like pSTOP1622 with additional sequence for N-terminal fusion of StrepII-tag, cleavable with TEV (tobacco etch virus) protease. pN-Strep-Xa1622 Like pSTOP1622 with additional sequence for N-terminal fusion of StrepII-tag, cleavable with Factor Xa protease. Vectors for special requirements pMGBm19 pMGBm19 is an E.coli/ Bacillus shuttle vector with xylose-inducible promoter (PxylA) that is designed for co-expression studies. It can be used in combination with any other vector of the 1520, 1622, and 1623hp series, since it contains an origin of replication of a different compatibility group (pMB100 replicon). pMMEc4 Since the xylose-inducible PxylA promoter is not tightly controlled in E. coli, cloning the toxic genes into vectors of the 1520, 1622, and 1623hp series, respectively, using E. coli as host may be difficult. In such cases, we recommend using the pMMEc4 helper plasmid. This E. coli vector (not replicating in Bacillus!) encodes xylose repressor XylR and is designed for blocking any expression starting from the PxylA promoter while cloning gene of interest within E. coli (Jordan et al., 2007). In pMMEc4 the expression of xylR is controlled by the arabinose-dependent promoter PBAD and the AraC protein. In the presence of 0.2% arabinose, the AraC protein binds to the operator sequence that activates the expression of the xylR gene, and additionally upregulates its own expression. The vector pMMEc4 carries the p15A origin of replication that is compatible with vectors from other incompatibility groups such as ColE1. Control vectors Vectors indicated below in the table are available as positive controls for expression in B. megaterium as well as for one-step affinity purification (except for pGFP1522). The vectors encode the GFP and different mutant proteins of levansucrase, an enzyme of Lactobacillus reuteri.
s three different B. megaterium strains (WH320, MS941, and YYBm1) for protein production. All strains are supplied as protoplasts, ready-to-use for transformation. 1. The strain WH320 is a chemical mutant of strain DSM319, which is deficient in the production of -galactosidase (lacZ). It was described by Rygus and Hillen (1991). 2. The strain YYBm1 carries the nprM deletion and an additional deletion of the xylose isomerase gene xylA. It is thus unable to metabolize xylose, which is used as inducer for gene activation (Yang et al. 2006). 3. The strain MS941 was generated from the wild-type strain DSM319 by deletion of major extracellular protease gene nprM (Wittchen and Meinhardt 1995). Because of reduced extracellular protease activity, this strain is well suited for extracellular protein production.

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