Process of Cloning

Molecular Cloning is one way of studying the specific proteins involved in cell division. A gene contains the instructions for how to make a protein. By mutating a gene, the protein’s shape, size and function could all be affected. Mutating a gene changes its instructions. Once a mutated gene is created and incorporated into a cell’s DNA, the cell replicates, creating many cells containing the mutant gene. The cells with the changed gene can then be compared to normal cells.

Below are the steps involved to both make a mutant gene and incorporate it into the DNA of a human cell:

  1. Chemically "cut" the gene you want to study from the DNA strand

    Cutting DNA for Target Gene
  2. Attach target gene to a small, circular piece of DNA.Together, this is called a plasmid, which serves as the vehicle for transporting the gene.

    Resulting Plasmid
  3. Put the plasmid into an E. coli cell (or another type of bacteria). As each E. coli cell divides, each new cell contains a copy of the plasmid containing the gene.

    Plasmid in E. Coli

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  1. Grow a lot of E. coli cells

    Lots of E. Coli
  2. Once your E. coli population has reached your desired number of cells, break apart the E. Coli cells using a chemical that dissolves the cell wall.

    Broken E. Coli Cells
  3. Filter the mixture of broken E. coli cells and collect only the plasmids containing the gene.

    Leftover Plasmids
  4. Put the plasmids into human cells. The type of cell varies depending on the research.

    Human Cell Containing Plasmid
  5. Over time, the plasmid will be incorporated into the host cell DNA and the new gene will change the proteins produced.

    Plasmid Incorporated into Human Cell
  6. Observe physical changes between the cells with the plasmid and those without.

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