Question

Some sources believe that azithromycin may hinder Covid infection by interfering with the E protein. You decide you want to make a coronavirus without an E protein to see if it is "attenuated", causing less dangerous infection, which might (with much trepidation) be used as a "contagious vaccine" [like cowpox]. In order to do this, you want to
a) convert the RNA genome of a coronavirus into a DNA copy
b) make lots of the DNA
c) either mutate that DNA or chop out a piece of it
d) produce RNA based on this DNA.
Suggest one or more biotechnology methods you might use for each of these steps.

Answer 1

a) To convert the RNA genome of a coronavirus into a DNA copy: ReverseTrascriptase enzyme is used to convert the single stranded RNA genome into complementary DNA (cDNA) .

b) To make lots of the DNA: Reverse trascriptase synthesized cDNA in step a) is used to amplify the lots of DNA by RT-PCR technique.

c) Either mutate that DNA or chop out a piece of it: Amplified DNA in step b) is used for site directed mutagenesis PCR and mutated DNA copies are produced  and Restriction enzymes (Restriction Endonucleases ) are used to cut the DNA in a pieces.

d) To produce RNA based on this DNA.: DNA product from step c) is used for trascription or synthesis of Interest of RNA by using DNA-dependent RNA- polymerase.This biotechnology method can be used for the Contagious Vaccine production.