Transforming Eukaryotic Cells

by Georgina Cornwall, PhD

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    00:00 So very exciting room for genetic technologies or biotechnologies. As I had introduced earlier, eukaryotic cells can be a little bit more challenging because they don't really want to transform. A number of different mechanisms have to be used. First of all, we tried the shotgun method and it literally is sort of short gunish. They take a gun and fire DNA literally at cells and hope that the cells take it in it. It is not particularly specific, but DNA will stick to gold or tungsten particles and they fire them at high velocity into the cells and just hump that they take up the DNA and that has proved to be fairly successful, but as I said not very directed. We are starting to come upon much more directed mechanisms that are really really cool. I will introduce you to one of them at the end of this lecture.

    00:53 Again, we have got a plasmid inside a bacterial cell. This particular plasmid is a Ti plasmid or a tumor inducing plasmid. We can use these tumor inducing plasmids to transmit information and see that the information was transmitted or DNA of interest. Tumor-inducing plasmids are in bacterial cells that often infect certain types of plants. When the bacterial cell infects the plant often the plasmid is transferred to the plant and induces a tumor and the plant will grow that tumor. So, what we can do is take these Ti plasmids, tumor inducing plasmids and use our restriction endonucleases to cut genes of interest, then stick them into that plasmid.

    01:43 And then when that plasmid is introduced to the plant cells, the plasmid can make its way into the plant cell nucleus because that is the point the bacteria was trying to infect the plant and so it tried to give it some DNA to mess things up and make a tumor and then those plant cells will replicate and the tumor grows and so we have a plant with this tumor on it. We can tell that the DNA of interest that we inserted into that bacterial plasmid has made it into the plant because it will develop a tumor. Or might not develop the tumor, but it depends on where the gene is inserted into the plasmid and how the plasmid makes its way into the plant. But we can get information genetic modifications in the plants using this mechanism.

    About the Lecture

    The lecture Transforming Eukaryotic Cells by Georgina Cornwall, PhD is from the course Biotechnology.

    Included Quiz Questions

    1. BAC ----- Transfer platinum particles coated with DNA to the animal cell
    2. Ti plasmid ----- Induce crown galls in Agrobacterium tumefaciens infected plants
    3. Biolistic particle delivery system ----- Transfer gold or tungsten particles coated with DNA to the host cell
    4. Agrobacterium ----- Natural genetic engineer
    5. Microinjection ----- Physical method of gene transfer in eukaryotic cells

    Author of lecture Transforming Eukaryotic Cells

     Georgina Cornwall, PhD

    Georgina Cornwall, PhD

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