Question

2. (Multiple answers) A student is doing an experiment to insert the plasmid seen below into an E. coli bacteria. They cut their plasmid and gene of interest with the HIND III restriction enzyme to insert their gene of interest. lacz HindIII BamHI -EcoRI amp PUC19 After the experiment is complete they observe many blue colonies on their plate media that contained nutrients, X-gal and ampicillin. Please choose all that are TRUE about this experiment. ampicillin is the selective ingredient in the media X-gal is being broke down by this organism This experiment successfully inserted the gene of interest into the plasmid The plasmid was taken up by the E.coli The E.coli can now produce the HIND III restriction enzyme.

Answer 1

Let's go through each statement and see whether it is true or not:

Statement 1: In the above experiment, the student wants to insert the given plasmid inside E. coli. In order to select the cells that have undergone successful transformations the vectors generally has an ampicillin resistance gene (ampR) and the plate where they are plated contains ampicillin as the selective ingredient. Thus the first statement is TRUE.

Statement 2: X-gal is broken down by beta-galactosidase. This process gives the blue color to the colonies. Beta-galactosidase is produced by lacZ gene. Since there are blue colonies present on the selective media we can say that the X-gal is being broken down. Thus the second statement is TRUE.

Statement 3: Since we are seeing blue colonies on the ampicillin + X-gal plate, we know that the plasmid vector has been successfully transformed. The blue color implies that the X-gal is being broken down which confirms the presence of lacZ gene. Thus the third statement is TRUE.

Statement 4: Due to the reasons given for the statement 3, the fourth statement is also TRUE.

Statement 5: The plasmid has the restriction site for HIND III, it can produce HIND III enzyme itself. Thus the fifth statement is FALSE.

Thus the correct statements are 1, 2, 3 and 4.

Hope this helped! All the best!