The helper phage vector was perfectly constructed for recombination, so why we have to use secondary plasmid like pLS7 or pNJB7 (Sattar et al., 2015- Ff-nano, Short Functionalized Nanorods Derived from Ff (f1, fd or M13) Filamentous Bacteriophage). So I have doubt why they not inserted the Ori1+PS+Ori2 in helper phage vector?. If I insert the Ori1+PS+Ori2 (~200bp) region into helper phage vector, then this will produce microphage or not. Please anyone explain me this, if it works then why they used separate plasmid for the above mentioned research article.
I'm not entirely sure that I understand your question, but I think you are asking why can you not clone an insert into a normal phage and generate microphage? I think the answer is that if you were to do so then the helper would no longer be able to fully replicate because of interference from the origins. In the helper phage scenario you have a plasmid that is acting as a template for microphage production but normal helper phage to provide replication and packaging proteins. Were you to interfere with the helper phage ability to function then you would be likely to get no product.
Thank you for your valuable answer Dr. Michael J. Benedik, yes sir the microphage origin will affect the helper phage replication but when the separate microphage origin plasmid transformed into E.coli it wont affect helper phage replication. I want to do microphage peptide display study, so I dont want the full helper phage. So in phagemid vector i can insert the microphage origin (-) instead of macrophage origin (+ & -), then I will get only microphage with peptide display. I have doubt on this will work or not. I will get only microphage with peptide display without helper phage infection. There is any possibility to get only microphage sir.
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