Amid Biosciences’ pTrham vector produces the highest yields of recombinant proteins in comparison to other rhamnose expression vectors available on the market. To ensure highly efficient transcription and translation of the cloned gene, L-rhamnose-inducible rhaBAD promoter of Escherichia coli is complemented with the rrnG anti-termination region, the bacteriophage T7 gene 10 translation enhancer and ribosome binding site.
This plasmid-based expression system with optimized transcription and translation regulatory elements is capable of efficient protein production upon induction with L-rhamnose but exhibits very low basal levels of expression in the absence of inducer. pTrham expression is tightly regulated at the individual cell level and promotes the high-level production of heterologous proteins in a correct and bioactive conformation. As a result, pTrham vector is capable of expressing a wide variety of recombinant proteins, including the stable cloning and expression of toxic proteins.
Small size of the pTrham vector (3.3 kb) facilitates cloning of large inserts and performing DNA manipulations, such as site-directed mutagenesis. The multiple cloning site (MCS) provides directional cloning options using a variety of restriction enzymes.
The pTrham expression vector is compatible with any E. coli strain.
Catalog # PTR-401
Figure 1. Analysis of Red Fluorescent Protein (RFP) expression by SDS-PAGE.
The pTrham vector containing a gene encoding RFP was transformed into BL21 (DE3) cells. Cells were grown to OD600 of 0.4 and induced with L-rhamnose (0.2%). Two parallel expressions were performed. Aliquots of cell culture were taken after 1, 2, 4 and 6 hours of induction and analyzed by SDS-PAGE (12 %). The gel stained with Coomassie ProStain shows RFP expression levels at different time points after rhamnose addition. M - protein markers; lanes 1, 2, and 7 - uninduced samples; 3 and 8 - one hour induction; 4 and 9 - two hours induction; 5 and 10 - four hours induction; 6 and 11 - six hours induction with rhamnose. No leaky expression can be seen in uninduced samples (lanes 1, 2, and 7).
Storage: -20 oC
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