Instructions for Use of Product(s)
Literature # 9PIN149
The pNLCoI4[luc2-P2A-NlucP/PGK/Hygro] Vector is a second generation coincidence reporter vector (CoI) that allows expression of both firefly luciferase (Fluc, luc2 gene) from Photinus pyralis and NanoLuc® Luciferase fused to a PEST destabilization domain (NanoLuc®-PEST, NlucP gene) from the same mRNA transcript. The stoichiometric expression of both luciferases is achieved by use of the P2A sequence from porcine teschovirus-1, which promotes a ribosomal skip and expression of the two unfused enzymes with different compound interaction profiles. When used in highthroughput screening, true hits will show a similar response from both reporters, reducing the number of false hits and the workload associated with follow-up screens.
The luc2 and NlucP genes are codon optimized for mammalian expression and all pNL vectors have minimal consensus transcription factor-binding sites to reduce anomalous expression. The pNLCoI4[luc2-P2A-NlucP/PGK/Hygro] Vector contains a PGK promoter for moderate-level expression of both reporters, allowing its use as a constitutive expression control.
Printed in USA. Revised 9/16