If I have to make the cloning strategy, what should be my workflow to do this.
I got two problems, one to select suitable expression vector from pCAMBIA1301, and pCAMBIA1302. As 1301 has GUS reporter protein and 1302 has mgfp5 (GFP), so what should be preferred.
The Other thing is for these vector we have to design a transgene cassette in MCS region. But the problem is how should I design my primers and what elements should be added in the transgene cassette.
And the second problem is how can we analyze our primer that it should work?
GUS is more accurate for identifying where the protein is expressed. This is only a concern if you are expressing your protein using the native promoter. As the pCAMBIA plasmids use the cauliflower mosaic virus promoter you probably want to go with 1302 for easier screening. Your primers should include the start codon and not include the stop codon. On the ends of your primers have separate restriction sites followed by 2-4bp of template sequence to allow for directional cloning into the MCS. Make sure you will be ligating in frame with the marker gene. You may wish to subclone into a TOPO vector. Use primer BLAST to check for predicted target binding and any off target binding sites. use cDNA to clone your GOI.
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