What is the name of the molecule that releases the 6-his tag from the column? Wh

Question

What is the name of the molecule that releases the 6-his tag from the column? What is the function of IPTG in the bacterial expression system we are using to express the 6-his:GFP protein? a. It causes the bacteria to divide more quickly, thereby producing more 6-his:GFP protein. b. It directly causes gene expression of the 6-his:GFP gene, by binding the laci and removing it from the operator upstream of the gene. c. It is a sigma factor for the 17 RNA polymerase. d. It causes gene expression of T7 RNA polymerase and of the 6-his:GFP gene by binding the laci and removing it from the operator upstream of both genes. When you put your four samples with 2 times SDS loading buffer in the freezer, the final volume of each sample should be a. 600 ul b. 25 ul c. 100 ul d. 50 ul

Explanation / Answer

Answer:

7. Imidazole

Imidazole competes with the his-tag for the Ni-binding sites in the beads, thus releasing the his-tagged proteins.

8. (b) It directly cause gene expression of the 6 his-GFP gene by binding the lacI and removing it from the operator upstream of the gene.

9. (c) 100 microL

50 ml of a column fraction should be combined with 50 ml of SDS PAGE 2x loading buffer, heated to 95°C for 3 minutes and then stored.

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