Which of the following experiments would be most useful to determine if the move

Question

Which of the following experiments would be most useful to determine if the movement of the intracellular bacterium Shigella inside its host cell depends on an actin tail (similar to Listeria).

Please note:

1. ShiA is a cell surface protein of Shigella.

2. As a result of Shigella movement inside the host cell, individual bacteria are distributed throughout the cytosol.

3. Secondary antibodies are not listed, but you can assume that the correct ones would be used to detect the primary antibody.

Express GFP-tagged actin and follow the movement of Shigella by live-imaging

Step1: block F-actin assembly by expression of large amounts of G-actin ATP sequestering protein
Step 2: fix cells and stain with an antibody to actin

Step1: block F-actin assembly by expression of large amounts of G-actin ATP sequestering protein
Step 2: fix cells and stain with an antibody to ShiA to determine the intracellular localization of the bacterium

Step 1: express a dominant negative form of myosin
Step 2: fix cells and stain them with an antibody to the bacterial protein ShiA to examine its intracellular localization

Fix cells and co-stain them with an antibody to the bacterial protein ShiA and an antibody to actin

Explanation / Answer

Express GFP-tagged actin and follow the movement of Shigella by live-imaging

Step1: block F-actin assembly by expression of large amounts of G-actin ATP sequestering protein

Step 2: fix cells and stain with an antibody to ShiA to determine the intracellular localization of the bacterium

Fix cells and co-stain them with an antibody to the bacterial protein ShiA and an antibody to actin

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