Protein Expression in E. coli using

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Protein Expression in E. coli using pEX
Expression Vectors

• Validation Data

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5 New E. coli Expression Vectors
Please see vector maps at http//www.origene.com/c
dna/TrueORF/DestinationVector.aspx then click on
Bacterial expression tab
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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.
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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

1. pEX-C-His-Bin1 (NM_139351) no IPTG
2. pEX-C-His-Bin1 (NM_139351) with IPTG

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Highly Efficient Protein Expression
pEX-N-His-GST vector shows comparabe high
expression level of GST with pGEX-2T vector.
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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).
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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.
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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.

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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.