Protein Expression Facility

pLTE vectors

The pLTE expression vectors are designed to be used in conjunction with the Leishmania tarentolae extract (LTE) in vitro translation system.

PEF offers 5 different vectors:

  1. pLTE (EGFP control)
  2. pLTE_6H_EGFP_3C
  3. pLTE_3C_EGFP_6H
  4. pLTE_6H_N and
  5. pLTE_6H_C

pLTE (EGFP control), pLTE_6H_EGFP_3C and pLTE_3C_EGFP contain an EGFP gene, which can be used as a positive control for transcription/translation reactions. pLTE_6H_N and pLTE_6H_C do not contain an EGFP gene. All vectors except the pLTE (EGFP control) contain a 6xHis-tag.

Additionally, all the vectors have a species independent translational sequence (SITS) incorporated upstream of the EGFP ORF. SITS consists of a poly(UUUUA)13 5′ UTR fused to a sequence which forms three mRNA stem-loop structures (Mureev et al., 2009). Translation of this sequence results in 17 additional amino acids at the N’ terminus of the target protein. This configuration ensures the highest yield of recombinant protein synthesis in the LTE and all other cell free expression systems.

You can choose to clone the gene of interest into the pLTE (EGFP control) vector via three cloning strategies:

  1. Replacement of EGFP control gene by target gene
  2. Fusion of target gene to N’ of EGFP gene
  3. Fusion of target gene to C’ of EGFP gene