| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | All primers used to modify pRP vectors are listed in Table S4. To integrate the firefly luciferase gene into the rRNA array, Fluc was amplified from pCRm-luc2-BSD and cloned into the SfiI sites of pRP-LA and pRP-VH (22). Insertion of the LdNT3 stem-loop into pRP-LA and pRP-VH was accomplished in a step-wise fashion. First, the vectors were subjected to whole-plasmid amplification to introduce two nonidentical BstXI sites into their shared 3’-UTRs (see primer sequences for detail). A version of the LdNT3 stem-loop was synthesized with flanking BstXI and PCR primer binding sites (Genscript, Piscataway, NJ) and inserted into the BstXI sites of the modified pRP vector. | Get A Quote |
The ability to modulate gene expression in response to changes in the host environment is essential for survival of the kinetoplastid parasite Leishmania. Unlike most eukaryotes, gene expression in kinetoplastids is predominately regulated post-transcriptionally. Consequently, RNA-binding proteins (RBPs) and mRNA-encoded sequence elements serve as primary determinants of gene regulation in these organisms; however, few have been ascribed roles in specific stress-response pathways. Leishmania lack the capacity for de novo purine synthesis and must scavenge these essential nutrients from the host. Leishmania have evolved a robust stress response to withstand sustained periods of purine scarcity during thei... More
