Transcript shRNA Customer Presentation

Protein Expression in E. coli Through
Various Destination Vectors
Validation Data
5 New E. coli Expression Vectors
pEX-Series of Destination Vectors for
Protein Expression in E. coli
ORF Expression in E. coli
E. Coli was transformed with 4
constructs with mammalian ORFs
in the pEX-N-His-GST vector.
GST-fused proteins were
expressed at high levels with all
constructs.
Highly Efficient Protein Expression
GE’s pGEX-2T bacterial expression
vector was compared with pEX-NHis-GST for protein expression
efficiency. Upon IPTG induction, two
vectors show comparably high
expression level of GST.
The Fused 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
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
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 cutter:
Sgf I and Mlu I or other specified
enzymes.
After a ligation reaction, the
resulting clones are grown on
ampicillin-containing medium to
select for successful subcloning of
the ORF into the destination
vector.
Order TrueORF in E. coli Vectors
For all destination vectors, there are two options:
1. Purchase both the TrueORF in pCMV6-Entry vector
and the desired destination vector and follow the
simple shuttling protocol.
2. Ask OriGene experts to perform the shuttling.
OriGene will provide highly purified ready-to-use
plasmids.