For the past 20 years many lab researchers will tell you that day-to-day ‘cloning’ is perhaps the most frustrating part of the work they do! However more recently, help has been at hand for those researchers with the emergence of many new cloning strategies that do not rely on conventional restriction enzyme digestions! Gateway cloning is one such method and speaking personally it certainly made things much easier in the lab (although perhaps not as simple as the designers might have you believe)!
Plant Physiology recently published an OA manuscript entitled ‘MultiSite Gateway-Compatible Cell Type-Specific Gene-Inducible System for Plants‘. This paper introduces a set of Gatway compatible constructs that combine plant selection markers, control of expression domains, access to multiple promoters and protein fusion reporters, chemical induction, and high-throughput cloning capabilities.
This ambitious goal was been acheived by mining research literature to select >20 promotors that provide tissue-type specific expression in the Arabidopsis root. These promotors have been linked with the estrogen-inducible XVE system which, again from personal experience, offers strong expression in response to an inducible signal. Therefore this will allow induced expression of a researchers gene of interest (GOI) in a specific cell file.
These constructs can be linked to a range of fluorescent proteins to allow visualisation of different tagged proteins in the same root as well as providing the facility to make both transcriptional or translational fusions.
In the paper the authors have tested the expression of many of these reporters and the whole set of clones are available either from the lab of Ari Pekka Mähönen (free) or from Addgene (pay). They look to be a very useful resource so please give them a go!
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