8. Suppose you have started to work in a virology laboratory and your advisor su

Question

8. Suppose you have started to work in a virology laboratory and your advisor suggests that you begin to study the function of Herpesvirus protein X. Answer the following questions 1:31 AM OO Sprint F 92% K Back Take-home Quiz 2.doc a) 2p. How could you use marker transfer techniques discussed in class to create a virus that was deleted for the protein X gene (gene X)? Explain how you would perform the experiment and identify your mutant virus, if gene X is essential for virus growth. b) 2p. You suspect that protein X is important for Herpesvirus DNA replication. What method would you use to determine if your protein X mutant is defective for producing normal amounts of viral DNA? Briefly explain how the method you choose works and why the method will measure amount of viral DNA. c)2p. You would now like to confirm the phenotype of your deletion mutant by making a less disruptive mutation that only changes the start codon of protein X so that it will no longer be recognized and translated by the ribosome. Assume that you have a cloned gene X. What mutagenesis procedure will you use to mutate the ATG start codon in your construct? Explain how the technique produces the desired mutation. A Courses Calendar To Do Notifications Messages

Explanation / Answer

1. To identify mutant viruses , we can perform mass spectrometry and also by protein mass mapping. Before the virus had been mutated a mass mapping study of the wild viral form along with a comparison of the mutated ( after treatment) viral form would help us to know if the virus is mutated.

In a laboratory, both the viral forms can be grown on a cell line. The wild form would be able to grow as it contains the protein X gene, the mutated form would not be able to grow and replicate as the protein X gene is absent in the viral genome .

2. Quantitation of the viral DNA can be done using a real time PCR of both non mutated and mutated viral genome , followed by amplification of the genome , internal standardisation followed by detection of the signals using a two colour florouscence detection , thus correctly estimating the of viral DNA .

3. A point mutation has to be produced in order to change the codon from U --->T , thus resulting a changed start codon from AUG TO ATG, which would disrupt the start codon of the X protein. This transition mutation can be brought about my chemical mutagenesis that would change or transit the codon. Alternatively, it can also be done using UV radiation.

Related Questions

Navigate

• Browse (All)
• Browse 8
• Subjects

---

---

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.