6. In an in vitro translation assay, radioactive amino acids can be incorporated

Question

6. In an in vitro translation assay, radioactive amino acids can be incorporated into proteins and the proteins separated from the unincorporated amino acids (higher counts per minute indicates more protein synthesis). Given the following data what are the conclusions that you would come to? RNA added to reaction Insulin mRNA alone Insulin mRNA with antisense insulin mRNA Insulin mRNA with antisense fibronectin RNA No mRNA Insulin mRNA with double stranded insulin RNA Amount of radioactivity in protein in counts per minute 10,000 3,000 11,000 150 500

Explanation / Answer

Please find the answers below:

The following conclusions can be drawn from the information given in the tabulated data:

Case 1: The insulin mRNA is usually translated into the desired protein under normal conditions.

Case 2: Anti-sense mRNA prevents free translation of the insulin mRNA, thus down-regulating total protein synthesis.

Case 3: Antisense fibronectin RNA does not base pair with insulin mRNA, thus translation takes place without any hinderance.

Case 4: Absence of mRNA deciphers altogether absence of translation.

Case 5: Since the mRNA is double stranded, it is not available for the access of translational machinery, hence the rate of protein synthesis is reduced.

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