not having expression in 293T cells

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bralmeidapt
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not having expression in 293T cells

Post by bralmeidapt » Fri Oct 28, 2005 3:00 pm

I have a x protein in a pCMV-Myc plasmid that sub-cloned in a pMSCV-IRES-GFP (that had a protein y) through EcoRI and XhoI for retroviral infection. Running a gel after digestion, looks that everything's alright and I transfected the pMSCV-y-IRES-GFP into 293T cells (as control) and pMSCV-x-IRES-GFP also into same cells both using Lipofectamine 2000.
After 48h I have expression of GFP in cells with protein y (control) but not in cells with protein x... interference from x mRNA with IRES structure??

Anyone has a clue?... :?

Thnks!

LaNcE222
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Post by LaNcE222 » Mon Nov 07, 2005 7:08 pm

WOW very interesting maybe they have a defense? Most likey not but it was a guess

bralmeidapt
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protection

Post by bralmeidapt » Tue Nov 08, 2005 9:04 am

Hum... that would probably not be the case - in theory, a cell transfected with a plasmid with a eucariotic promoter (the case of pMSCV) can process any protein that you insert... and if that would be the case, protein y wouldn't also be expressed :wink:

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