Synthesis of the amino acid histidine is a multistep anabolic pathway that uses the products of 13 genes (hisA to hisM) in E. coli. Two independently isolated  E. coli mutants, designated his1⁻ and his2⁻ are studied in a conjugation experiment. A his⁺ F' donor strain that carries a copy of the hisJ gene on the plasmid is mated with a his1⁻ recipient strain in Experiment 1 and with a his2⁻ recipient in Experiment 2. The exconjugants are grown on plates lacking histidine. Growth is observed among the exconjugants of Experiment 2 but not among those of Experiment 1.

Why is growth observed in Experiment 2 but not in Experiment 1?

An Hfr strain with the genotype cys⁺ lue⁺ met⁺ strˢ is mated with an  strain carrying the genotype cys⁻ lue⁻ met⁻ strᴿ. In an interrupted mating experiment, small samples of the conjugating bacteria are withdrawn every 3 minutes for 30 minutes. The withdrawn cells are shaken vigorously to stop conjugation and then placed on three different selection media, composed as follows:

Medium 1: Minimal medium plus leucine, methionine, and streptomycin
Medium 2: Minimal medium plus cysteine, methionine, and streptomycin
Medium 3: Minimal medium plus cysteine, leucine, and streptomycin

What is the purpose of adding streptomycin to each selection medium?

An Hfr strain with the genotype cys⁺ lue⁺ met⁺ strˢ is mated with an  strain carrying the genotype cys⁻ lue⁻ met⁻ strᴿ. In an interrupted mating experiment, small samples of the conjugating bacteria are withdrawn every 3 minutes for 30 minutes. The withdrawn cells are shaken vigorously to stop conjugation and then placed on three different selection media, composed as follows:

Medium 1: Minimal medium plus leucine, methionine, and streptomycin
Medium 2: Minimal medium plus cysteine, methionine, and streptomycin
Medium 3: Minimal medium plus cysteine, leucine, and streptomycin

List all possible bacterial genotypes growing on each medium.

An Hfr strain with the genotype cys⁺ lue⁺ met⁺ strˢ is mated with an  strain carrying the genotype cys⁻ lue⁻ met⁻ strᴿ. In an interrupted mating experiment, small samples of the conjugating bacteria are withdrawn every 3 minutes for 30 minutes. The withdrawn cells are shaken vigorously to stop conjugation and then placed on three different selection media, composed as follows:

Medium 1: Minimal medium plus leucine, methionine, and streptomycin
Medium 2: Minimal medium plus cysteine, methionine, and streptomycin
Medium 3: Minimal medium plus cysteine, leucine, and streptomycin

What donor gene is the selected marker in each medium?

Synthesis of the amino acid histidine is a multistep anabolic pathway that uses the products of 13 genes (hisA to hisM) in E. coli. Two independently isolated  E. coli mutants, designated his1⁻ and his2⁻ are studied in a conjugation experiment. A his⁺ F' donor strain that carries a copy of the hisJ gene on the plasmid is mated with a his1⁻ recipient strain in Experiment 1 and with a his2⁻ recipient in Experiment 2. The exconjugants are grown on plates lacking histidine. Growth is observed among the exconjugants of Experiment 2 but not among those of Experiment 1.

What is the genotype of exconjugants in Experiment 2?

Look closely at the consolidated Hfr map and the data used to build the map on page 261. Suppose a fifth Hfr strain had the F factor inserted exactly halfway between cysE and leuU and had an orientation that was the same as that of Hfr 1. List the order of gene transfer for the first six genes transferred by this Hfr and the number of minutes of conjugation at which each gene is expected to be seen.