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A series of dual-reporter vectors for ratiometric analysis of protein abundance in plants

biorxiv. 2020; 
Aashima Khosla,  Cecilia Rodriguez-Furlan,  Suraj Kapoor,  Jaimie M. Van Norman,  David C. Nelson
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Molecular Biology Reagents The plant codon optimized coding sequence of mScarlet-I, LUC2, mCeruelan-NLS, and redLUC-*F2A-gLUC were synthesized in pUC57 cloning vectors (Genscript) using KpnI - SacI, ApaI-MluI, KpnI-SacI, and MluI-SacI restriction sites, respectively. Get A Quote

摘要

Ratiometric reporter systems enable comparisons of the abundance of a protein of interest, or “target,” relative to a reference protein. Both proteins are encoded on a single transcript but are separated during translation. This arrangement bypasses the potential for discordant expression that can arise when the target and reference proteins are encoded by separate genes. We generated a set of 18 Gateway-compatible vectors termed pRATIO that combine a variety of promoters, fluorescent and bioluminescent reporters, and 2A “self-cleaving” peptides. These constructs are easily modified to produce additional combinations or introduce new reporter proteins. We found that mScarlet-I provides the best signal-t... More

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