Protein Expression in E. coli Through Various Destination Vectors

1. Protein Expression in E. coli Through Various Destination Vectors Validation Data

2. 5 New E. coli Expression Vectors pEX-Series of Destination Vectors for Protein Expression in E. coli

3. 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.

4. Highly Efficient Protein Expression GE’s pGEX-2T bacterial expression vector was compared with pEX-N-His-GST for protein expression efficiency. Upon IPTG induction, two vectors show comparably high expression level of GST.

5. 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).

6. 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.

7. 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.

8. 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.