Mutated MDCK cells - protocol for mutating MDCK cells (Jul/23/2009 )
Hello everyone,
You might find this topic in the cell and tissue culture section. But I will brief you about the problem I am facing.
I am trying to express a viral protein in MDCK cells, mammalian cells. Well that is normal work. But to add spice to the work, I thought of mutating the MDCK cells, mammalian cells so that I can see the expression of the viral proteins via cloning in these cells.
I am desperately looking for a protocol which is very easy to follow (in the sense very easy to make cancer MDCK cells in the lab). It must be economical and easy to make. I have 2 weeks to finish this work.
Please help me.
Thanks you
Miracle Strain
miraclestrain on Jul 23 2009, 08:23 PM said:
You might find this topic in the cell and tissue culture section. But I will brief you about the problem I am facing.
I am trying to express a viral protein in MDCK cells, mammalian cells. Well that is normal work. But to add spice to the work, I thought of mutating the MDCK cells, mammalian cells so that I can see the expression of the viral proteins via cloning in these cells.
I am desperately looking for a protocol which is very easy to follow (in the sense very easy to make cancer MDCK cells in the lab). It must be economical and easy to make. I have 2 weeks to finish this work.
Please help me.
Thanks you
Miracle Strain
It depends upon what type of mutation you are looking for.
If you are looking for overexpressing a protein, or knocking down a gene, you just need to make a overexpression vector or shRNA vector and make a stable MDCK cell line.
If you are looking for directed mutagenesis, it is much tougher to achieve that in any mammalian cells other than mouse ES cells. There are protocols for doing that, however. Once you have clarified your project I can guide you in right direction.