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Expression GFP with pMDC83/85


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#1 aces

aces

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Posted 20 April 2011 - 02:37 PM

Hi all,

I am very new to gateway system. I want to express a 795 bp gene of interest tagged with GFP in fungus and I have inherited the plasmid pMDC83 and pMDC85. They contain different cassettes and different sizes of attR1 sequences.

pMDC83 (cassette A) .. TAT CAC AAG TTT GTA CAA AAA A
pMDC85 (cassette C) .. TAT CAC AAG TTT GTA CAA AAA AAA

I doubt which one fits with my entry clone produced with pENTR/D-TOPO? I guess from the attR1 sequence that the differences are about reading frame. Should I add two more bases into my forward to complete the reading frame if I use pMDC83?

Can anybody help me on this system? I tried reading up from invitrogen website but don't quite understand.

Thank you in advance.

/Ace




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