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Apple1984

Member Since 22 Jan 2013
Offline Last Active May 14 2013 01:34 PM

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2xzwei
21 Mar 2013 - 04:04
pito
22 Jan 2013 - 13:40

Topics I've Started

Packaging vectors

26 January 2013 - 08:23 AM

Hi,
Anybody knows the differences of D8.2 and D8.9?
I read a paper they used D8.9 for the tritransfection with lentiviral vector. But i have D8.2, instead of D8.9 and i am wondering if i can use D8.2.

Many thanks.

NW

lentivirus infection

22 January 2013 - 01:02 PM

Hi,
I am wondering if anybody has the same experience as me.
I generated lentivirus with a MSCV vector with GFP at the N terminal, encoding full length of my protein.
With the viral stocks, I infected multiple cancer cell lines. i could see green fluorescence under microscope and sorted the cells by flow cytometry. I did see lots of fluorescence after sorting. But the expression levels of the protein didn't change at all, tested by western blot.

I don't understand. If it is because of the cell toxicity of my protein.....? But why i could see so much fluorescence?
I am considering to switch another vector. But i still don't understand what happened to my protein. How could the GFP and my protein be separated?

If anybody can help me figure this out?

Many thanks.