Here I am talking in context with transgenic plants. For example can we get desired results on addition of Gus gene as a scorable marker and herbicide resistant gene as a selectable marker in single gene construct. This gene construct also contains gene of interest and its promoter.
Dear Divy Sharma ,
It is commonly done to increase the accuracy of the screening.
I recommend to check the vector using a restriction assay (extract dna, digest it and run an electrophoresis), so you corroborate that the screening's result is due to the promoter activity and not influenced by errors in the vector transformation.
Divy Sharma
Yes, you can. See this binary vector, which contains GUS, GFP screenable makers and hygromycin selectable marker (Hygr). Figure from: https://www.addgene.org/64401/
Divy Sharma
pCAMBIA1301.
Here is another popular Agrobacterium-mediated plant genetic transformation binary vector pCAMBIA1301. You can see that the construct contains GUS gene (a screenable marker), Hygromycin-resistant gene (Hygr, a selectable marker) and a multiple-cloning site (MCS) where you can place your gene-of-interest (GOI) into it.
Yes this is quite possible and a better way if getting true transgenes.
Divy Sharma
By the way, you said "Can a single gene construct carry selectable and scorable/ reporter marker together?" 👉 It is better to say 'a single T-DNA construct', instead of 'a single gene construct'. These genes are within two T-DNA borders, the LB and RB.
All depends on what you want to do and which promoters you are using.
You can think also of using "2A" system to link multiple genes under the control of a single promoter...etc. See Article Self-processing 2A-polyproteins - A system for co-ordinate e...
andChapter Using the 2A Protein Coexpression System: Multicistronic 2A ...
In brief, yes, you can build/create/obtain such a vector. The pCAMBIA example is a classic one. These vectors do tend to be rather large, which can make genetic cloning a bit more challenging than smaller vectors.
Often vectors will contain just the selection marker and your GOI, without another transgene. One thing to keep in mind if you want to use GUS is that tissues are fixed (dead) during the staining protocol so you have to either wait until plantlets are large enough to sample from or sacrifice some plantlets for staining purposes.
Also check https://www.sciencedirect.com/topics/agricultural-and-biological-sciences/selectable-marker
Hello Sharma,
You can look at this paper.
Article Alternatives to Antibiotic Resistance Marker Genes for In Vi...
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