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Protein Expression in E. coli using pEX Expression Vectors. Validation Data. 5 New E. coli Expression Vectors. Please see vector maps at http://www.origene.com/cdna/TrueORF/DestinationVector.aspx then click on “Bacterial expression” tab . Protein Expression in E. coli N-tagged vector.

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Presentation Transcript
slide1

Protein Expression in E. coli using

pEX Expression Vectors

  • Validation Data
slide2

5 New E. coli Expression Vectors

Please see vector maps at http://www.origene.com/cdna/TrueORF/DestinationVector.aspx

then click on “Bacterial expression” tab

protein expression in e coli n tagged vector
Protein Expression in E. coli N-tagged vector

4 different mammalian ORFs was cloned into the pEX-N-His-GST vector.

GST-fused proteins were expressed at high levels with all constructs.

slide4

Protein Expression in pEX-C-His Vector

Protein Expression in pEX-C-His Vector

1 2

MW (kDa)

191

97

The ORF of Bin1 was cloned into

pEX-C-His vector and expressed in BL21(DE3) cells.

Total protein lysates were loaded on the SDS-PAGE gel.

64

BIN1

51

39

28

  • pEX-C-His-Bin1 (NM_139351) no IPTG
  • pEX-C-His-Bin1 (NM_139351) with IPTG
highly efficient protein expression
Highly Efficient Protein Expression

pEX-N-His-GST vector shows comparabe high expression level of GST with pGEX-2T vector.

the fusion tag can be cleaved
The Fusion Tag Can Be Cleaved

A purified GST fusion protein was incubated with TEV protease at 30oC for the indicated period of time. The recombinant protein and GST tag were separated by TEV cleavage (C).

pex 1 for untagged expression
pEX-1 for Untagged Expression

When no-tag is desired for the recombinant protein, pEX-1 is the vector of choice, appending only two amino acid (T and R) to the C-terminus of the protein.

easy trueorf shuttling into pex
Easy TrueORF Shuttling into pEX

37,000 TrueORF clones can be easily shuttled from entry vector into E. coli pEX vectors.

The entry and destination vectors are digested with two rare cutters: Sgf I and Mlu I or other specified enzymes.

For pEX-1 and pEX-C-His vectors use BseR I and MluI for subcloning.

*

order trueorf in e coli vectors
Order TrueORF in E. coli Vectors
  • For all destination vectors, there are two options:
  • Purchase both the TrueORF in pCMV6-Entry vector and the desired destination vector and follow the simple shuttling protocol.
  • Ask OriGene experts to perform the shuttling. OriGene will provide highly purified ready-to-use plasmids.