You have an Hfr strain of E. coli that contains wild-type genesfor maltose catabolism and alanine biosynthesis. You want totransfer these two genes into an F- strain that cannot use maltoseand is auxotrophic for alanine, by conjugation. Your Hfr strainalso has a tetracycline resistance gene located distal to theorigin of transfer. You have a P1 phage suspension and any ONEstrain of E. coli given in the MBIO 2020 lab manual at yourdisposal. Outline briefly how you would use these materials tocarry out a successful conjugation and transfer the two genes ofinterest. You do not need to include experimental details, but youshould give a medium that could be used to select for thesuccessful recombinants after each gene transfer that you use. Inthe box below, briefly answer the following questions:
Why can’t you just conjugate the two strains as they are? (ie.why do you even need the P1 phage to carry out thisexperiment?)
What ONE additional E. coli strain from the lab manual would youuse?
Outline a brief plan to transfer the mal and ala genes from theHfr strain to the F- strain. Do not include experimental details(reagents, incubation times, number of test tubes, etc). DO INCLUDEan appropriate selection step for every gene transfer experimentthat you use.
