I have identified micro RNAs and their potential targets boinformatically and now want to validate their interaction by coexpression system, in which, i want to clone target gene region under/at 3'UTR region of reporter (GUS/GFP) gene. So that if mature micro RNA will be interacting the target region the expression of reporter gene will be downregulated/off. Can anybody help me in finding any suitable viral vector for Agrobacterium mediated transient expression, in which there is possibility of inframe cloning of 100bp within or at 3UTR of the reporter gene. I want to use that vector for cucumber infiltration. If such vector are commercially available I can purchase it.
Hi Cheema,
Probably you can use TRV. Read the article doi: 10.1104/pp.113.231100. However, you will have to test first if TRV is systemic in cucumber.
The other possibility is to see if "Gateway-adapted expression vectors
containing validated microRNA (miRNA) sequences for use as positive controls" can be used in plants.
Hope it helped.
Good luck
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