Michael Rehli • Dept. Internal Medicine III • University Hospital • F.-J.-Strauss Allee 11 • 93053 Regensburg
fon: **49 (0)941 944 5587    fax: **49 (0)941 944 5593

Materials


Our group has generated and published a number of useful reagents that are available to researchers on request. If you wish to obtain any of the vectors, please state exactly which constructs you wish to obtain. If required, please complete the provided forms.
    Vectors
  1. pCpGL
  2. pMT-Bip/MBD-Fc
  3. pGL3-Toll-like receptor promoters
  4. pGL3-Chitinase promoters
    Cell-lines Coming soon!
  1. U937-pSwitch
  2. THP1-pSwitch
  3. S2-pMT-BIP/MBD-Fc
Vectors are ususally spotted on Whatman paper and sent via regular mail. Cell lines must be sent by courier mail either frozen on dry ice (pSwitch clones) or as liquid cultures (S2 cells). Please provide your favourite courier account number (E.g. FedEX or World Courier) and provide Customs declarations if required.






pCpGL vector  

pCpGL is a vector designed for reporter studies on the effect of promoter CpG methylation. The vector is completely CpG-free and its backbone does not influence the activity of the luciferase reporter gene after in vitro treatment with CpG methylases like SssI, HhaI or HpaII.

(Reference: Klug & Rehli (2006), Epigenomics 1, 127-130. [Abstract] )
pCpGL-basic (empty reporter vector, CpG-free)
pCpGL-CMV/EF1 (positive control, CpG-free)

Map and sequence information are available for download. [map], [seq]

The pCpGL plasmids contain the R6K gamma origin of replication and require special E.coli strains, like PIR1 (Invitrogen), GT115 (Invivogen). The plasmid also carries a ZEOCIN resistence gene.

Because INVIVOGEN sells the parental CpG-free vectors with a Limited Use License, this plasmid may be transfered to a scientific collaborator, provided that such transfer is not for any Commercial Purpose, and that such collaborator agrees in writing (a) not to transfer the vector or derivatives to any third party, and (b) to use the vector solely for research and not for Commercial Purposes. If you wish to obtain this vector, please sign the corresponding declaration and send it by FAX or email along with your request. Thank you.

Download the declaration form (pdf-file)

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pMT-BIP/MBD-Fc vector  

pMT-BIP/MBD-Fc is a metal-inducible expression vector for Drosophila S2 Schneider cells enabeling the expression of our proprietary MBD2-Fc fusion protein. This protein has high affinity for methylated CpG-DNA and can be used to detect methylation densities of genomic DNA fragments. The bivalent polypeptide and its applications are patend pending - transfer of this vector requires a signed Material Transfer Agreement.

(References: Gebhard et al. (2006) ,Cancer Res. 66, 6118-28. [Abstract] & Gebhard et al. (2006), Nucleic Acids Res. 34, e82. [Abstract])
pMT-BIP/MBD-Fc (expression vector for MBD-Fc polypeptide)


Because patents are pending for this plasmid it may be transfered to scientific collaborators, provided that the collaborator's institution signs the corresponding Material Transfer Aggreement (MTA). The vector or derivatives may not be transfered to any third party, and solely used for research and not for Commercial Purposes. To obtain the MTA, please download the request form and email the completed form along with your request.

Download the MTA request form (rtf-file)

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pGL3-based reporter vectors for Toll-like receptor genes  

Our lab has generated a large number of reporter plasmids for human and mouse Toll-like receptor genes. A list of constructs for each TLR gene available is given below.

TLR4 promoter
(Reference: Rehli et al. (2000), J. Biol. Chem. 275, 9773-81. [Abstract])


pGL3-TLR4–4121
pGL3-TLR4–2416
pGL3-TLR4–4121_Ex–3228-–432
pGL3-TLR4–742
pGL3-TLR4–385
pGL3-TLR4–75
pGL3-TLR4–75mO
(mutation of proximal OCT motif)
pGL3-TLR4–75mP1 (mutation of distal PU.1 motif)
pGL3-TLR4–75mP2 (mutation of proximal PU.1 motif)
pGL3-TLR4–75mI (mutation of IRF motif)
pGL3-TLR4–75mP12 (mutation of distal and proximal PU.1 motif)

(Reference: Lichtinger et al. (2007), J. Biol. Chem. 282, 26874-83. [Abstract])


pGL3-TLR4–385mP0 (mutation of PU.1 motif)
pGL3-TLR4–385mP1 (mutation of PU.1 motif)
pGL3-TLR4–385mP2 (mutation of PU.1 motif)
pGL3-TLR4–385mP5 (mutation of PU.1 motif)

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Tlr4 promoter (–428 - +112)
(Reference: Lichtinger et al. (2007), J. Biol. Chem. 282, 26874-83. [Abstract])


pGL3-Tlr4–428
pGL3-Tlr4–428_mP1
(mutation of PU.1 motif)
pGL3-Tlr4–428_mP2 (mutation of PU.1 motif)
pGL3-Tlr4–428_mP3 (mutation of PU.1 motif)
pGL3-Tlr4–428_mP4 (mutation of PU.1 motif)
pGL3-Tlr4–428_mP5 (mutation of PU.1 motif)
pGL3-Tlr4–428_mP6 (mutation of PU.1 motif)
pGL3-Tlr4–428_mP7 (mutation of PU.1 motif)
pGL3-Tlr4–428_mP8 (mutation of PU.1 motif)


Tlr4 promoter (–428 - –141)

pGL3-dTlr4–428
pGL3-dTlr4–428_mA/E
(mutation of AP1/E-Box motif)
pGL3-dTlr4–428_mP4 (mutation of PU.1 motif)
pGL3-dTlr4–428_mP5 (mutation of PU.1 motif)
pGL3-dTlr4–428_mP6 (mutation of PU.1 motif)
pGL3-dTlr4–428_mP7 (mutation of PU.1 motif)
pGL3-dTlr4–428_mP8 (mutation of PU.1 motif)

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TLR2 promoter
(Reference: Hähnel et al. (2002), J. Immunol. 168, 5629-37 [Abstract])


pGL3-TLR2–2750
pGL3-TLR2–2070
pGL3-TLR2–370
pGL3-TLR2–220
pGL3-TLR2–64
pGL3-TLR2–45
pGL3-TLR2+24
pGL3-TLR2–220_mS1
(mutation of proximal Sp1 motif)
pGL3-TLR2–220_mS2 (mutation of distal Sp1 motif)
pGL3-TLR2–220_mS12 (mutation of both Sp1 motifs)
pGL3-TLR2–64_mS1 (mutation of proximal Sp1 motif)
pGL3-TLR2–64_mE (mutation of E-box motif)
pGL3-TLR2–64_mES1 (mutation of E-box and Sp1 motifs)

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TLR3 promoter
(Reference: Heinz et al. (2003), J. Biol. Chem. 278, 21502-21509 [Abstract])


pGL3-TLR3–588
pGL3-TLR3–588_mI
(mutation of IRF motif)
pGL3-TLR3–588_mS (mutation of STAT motif)

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Tlr3 promoter
(Reference: Heinz et al. (2003), J. Biol. Chem. 278, 21502-21509 [Abstract])


pGL3-Tlr3–966
pGL3-Tlr3–966_mI-in
(mutation of inner IRF motif)
pGL3-Tlr3–966_mI-out (mutation of outer IRF motif)

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pGL3-based reporter vectors for Chitinase family genes  

A number of reporter plasmids for human CHI3L1 and CHIT1 promoters are available. A list of constructs for each gene is given below.


CHIT1 promoter
(Reference: Pham et al. (2007), J Biol Chem. 282, 21924-33. [Abstract]
)

pGL3-CHIT1–1503
pGL3-CHIT1– 419
pGL3-CHIT1–357
pGL3-CHIT1–247
pGL3-CHIT1–149
pGL3-CHIT1–76
pGL3-CHIT1–76mC
(mutation of proximal C/EBP motif)
pGL3-CHIT1–76mS (mutation of proximal GC box)


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CHI3L1 promoter
(Reference: Rehli et al. (2003), J Biol Chem. 278, 44058-67. [Abstract]
)

pGL3-CHI3L1–4400
pGL3-CHI3L1–3300
pGL3-CHI3L1–1300
pGL3-CHI3L1–1023
pGL3-CHI3L1–752
pGL3-CHI3L1–523
pGL3-CHI3L1–433
pGL3-CHI3L1–377
pGL3-CHI3L1–308
pGL3-CHI3L1–231
pGL3-CHI3L1–217
pGL3-CHI3L1–206
pGL3-CHI3L1–190
pGL3-CHI3L1–180
pGL3-CHI3L1–144
pGL3-CHI3L1–125
pGL3-CHI3L1–92
pGL3-CHI3L1–377_ExAC
(internal deletion)
pGL3-CHI3L1–377_ExSB (internal deletion)
pGL3-CHI3L1–377_ExZT (internal deletion)
pGL3-CHI3L1–377_mA (mutation of putative binding site A)
pGL3-CHI3L1–377_mC (mutation of putative binding site C)
pGL3-CHI3L1–377_mS (mutation of putative binding site S)
pGL3-CHI3L1–377_mB (mutation of putative binding site B)
pGL3-CHI3L1–377_mE (mutation of putative binding site E)
pGL3-CHI3L1–377_mZ (mutation of putative binding site Z)
pGL3-CHI3L1–377_mP (mutation of putative binding site P)
pGL3-CHI3L1–377_mCB (mutation of putative binding sites C and B)
pGL3-CHI3L1–377_mSB (mutation of putative binding sites S and B)
pGL3-CHI3L1–377_mSE (mutation of putative binding sites S and E)
pGL3-CHI3L1–377_mSP (mutation of putative binding sites S and P)
pGL3-CHI3L1–377_mSC (mutation of putative binding sites S and C)
pGL3-CHI3L1–377_mSBE (mutation of putative binding sites S, B and E)
pGL3-CHI3L1–377_mSBEP (mutation of putative binding sites S, B, E and P)
pGL3-CHI3L1–263_ExZT (internal deletion)
pGL3-CHI3L1–263_ExZT_mC (mutation of putative binding site C)
pGL3-CHI3L1–263_ExZT_mS (mutation of putative binding site S)
pGL3-CHI3L1–263_ExZT_mSC (mutation of putative binding sites S and C)


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