182-2010-ps5Key - 1 You isolated three T4 phage mutants...

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1. You isolated three T4 phage mutants from the rII region of the phage (1,2,3). When tested individually, the mutant phage form plaques on E. coli B but do not form plaques an E. coli K strain. You realize you can characterize these mutants using Benzer’s methods. You make pair-wise mixtures of the phage and use the mixtures to infect E. coli K at high MOI. You then ask if plaques form. 1 2 3 1 - + - 2 + - - 3 - - - What have you learned from these experiments about the three mutations? You then mixed the phage strains pair-wise and infect B line at high m.o.i. You allow enough time for the phage to lyse the cells and then determine the titre of phage by infecting E. coli K and E. coli B. You plot your data as (# plaques on K)/(# of plaques on B) . The ratios you obtain are as follows 1 2 3 1 1x10 -7 0.015 2x10 -7 2 3x10 -7 0.003 3 0 Please draw a map of the 3 markers and indicate the type (point mutation or deletion) and the positions of three mutations as well as the map distance between them. RF m1m2 = (2 x 0.015)/1.015 x 100 = 3.0 RF m2m3 = (2 x 0.003)/1.003 x 100 = 0.6 Other points to make: 1) first experiment is complementation test and second is recombinant assay;
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2) In comp. test, 1 and 2 can complement, so they are located in different
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