Question

Arrange the following steps in the correct order, outlining the transformation of bacteria using a plasmid encoding ampicillin resistance and green fluorescent protein (GFP). Assign 10 points to each step.

- Treat ampicillin-sensitive cells with electrical shock or exposure to calcium chloride to make them artificially competent.

- Introduce cells to plasmid encoding ampicillin resistance and fluorescence.

- Cell membrane becomes temporarily more permeable to DNA.

- Select cells positive for transformation.

- Plate cells on media containing ampicillin.

Answer 1

The correct order for transforming bacteria using a plasmid encoding ampicillin resistance and GFP is: treating cells, introducing cells to the plasmid, temporary permeabilization of the cell membrane, selecting positive cells, and plating cells on ampicillin media.

Step-by-step explanation:

The correct order of the steps for transforming bacteria using a plasmid encoding ampicillin resistance and green fluorescent protein (GFP) is as follows:

1. Treat ampicillin-sensitive cells with electrical shock or exposure to calcium chloride to make them artificially competent.
2. Introduce cells to plasmid encoding ampicillin resistance and fluorescence.
3. Cell membrane becomes temporarily more permeable to DNA.
4. Select cells positive for transformation.
5. Plate cells on media containing ampicillin.

This order ensures that the ampicillin-resistant and GFP-expressing cells are successfully transformed and selected. The treatment of cells with electrical shock or calcium chloride enhances the ability of the cells to take up the plasmid. The introduction of the plasmid provides the transformed cells with ampicillin resistance and GFP fluorescence. The selection process identifies the cells that have successfully taken up the plasmid and the plating on ampicillin media further confirms the presence of the ampicillin resistance gene.