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This Week. Score Conjugation Plates, Start High Frequency of Recombination (HFR) experiment, Continue Nasonia experiment. H igh F requency of R ecombination (Hfr). ...bacteria exhibiting a high frequency of recombination , - PowerPoint PPT Presentation
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This Week
• Score Conjugation Plates,
• Start High Frequency of Recombination (HFR) experiment,
• Continue Nasonia experiment.
High Frequency of Recombination(Hfr)
...bacteria exhibiting a high frequency of recombination,
– an alteration DNA sequence such that the genotype of subsequent individuals differs from the parent,
…specifically, strains with a chromosome integrated F factor that is able to mobilize and transfer part of the chromosome to the F- cell.
Hfr Cells
F factor
Bacterial Chromosome
Inserted F plasmid
...F factor integration site,
...host (bacteria chromosome) integration site.
F Pilus Attaches to F- Cell
Hfr DNA is Cut
F factor and Chromosomal DNA are Transferred
Recombination Requires Crossing over
Double Crossover
DNA not Incorporated into Chromosome are Digested
F factor inserts in different regions of the bacterial chromosome,
Also inserts in different orientations.
Origin of Replication
Hfr Order of transferstrain
H thr azi ton lac pur gal his gly thi 1 thr thi gly his gal pur lac ton azi 2 lac pur gal his gly thi thr azi ton 3 gal pur lac ton azi thr thi gly his
F factor
Hfr F-
A
a
Indicates direction of transfer.
A
A
a
Hfr DNA that is not incorporated in the F- strand, and DNA that has crossed out of the F- strand is
digested.
F factor
Hfr F-
A
Leading Gene: the first gene transferred is determined empirically.
A
Hfr
A
F-
A
A transfers first.
A transfers last.
Hfr Order of transferstrain
H thr azi ton lac pur gal his gly thi 1 thr thi gly his gal pur lac ton azi 2 lac pur gal his gly thi thr azi ton 3 gal pur lac ton azi thr thi gly his
E. coli Map
• 0 minutes is at the threonine,
• 100 minutes is required to transfer complete genome,