Standard recombination between an F’ and the chromosome.
-
Upload
rosa-cunningham -
Category
Documents
-
view
213 -
download
0
Transcript of Standard recombination between an F’ and the chromosome.
![Page 1: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/1.jpg)
Standard recombination between an F’ and the chromosome
![Page 2: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/2.jpg)
Recombination between an F’ and a strain with a deletion (this doesn’t
give Lac+)
![Page 3: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/3.jpg)
Recombination between an F’ and a strain with a deletion (this does give
Lac+)
![Page 4: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/4.jpg)
Jeffery Miller made a series of ordered
deletions of the lacI gene
Ach!! about 1 amino acid apart!
![Page 5: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/5.jpg)
He used these map mutations in lacI
![Page 6: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/6.jpg)
An example from Miller’s workSchmeissner_JMB_1977
Horizontal red lines show where the mutations must lie—between the endpoints ofthe deletions to their immediate left and right
![Page 7: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/7.jpg)
LacI monomer
![Page 8: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/8.jpg)
LacI dimer bound to lacO
![Page 9: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/9.jpg)
LacI dimer bound to DNA w/ HTH shown
![Page 10: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/10.jpg)
Close up of LacI HTH bound to DNA
![Page 11: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/11.jpg)
Binding of regulators to DNA is often at inverted repeats
Regulator proteins bind asback-to-back dimer. This puts each binding domainin the right orientation withrespect to the inverted repeat(protein toe binds 5’ end, heelbinds 3’ end of the repeat)
Binding siteshave invertedrepeats
Inverted repeats areon the same sideof the helix (spaced~10 bp apart)
![Page 12: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/12.jpg)
LacI dimer forms back to back
![Page 13: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/13.jpg)
Other Helix-Turn-Helix (HTH) DNA-binding proteins
TraR repressor Crp activator
![Page 14: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/14.jpg)
Close up of Crp HTH bound to lac DNA
![Page 15: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/15.jpg)
Mutations that give rise to stop codons
![Page 16: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/16.jpg)
Frequencies of lacI mutations generated by different mutagens
![Page 17: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/17.jpg)
Locations of nonsense mutations in LacI
![Page 18: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/18.jpg)
Locations of nonsense mutations in LacI
![Page 19: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/19.jpg)
Activity of suppressed nonsense mutations
![Page 20: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/20.jpg)
Recombination frequencies between closely linked nonsense mutations
![Page 21: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/21.jpg)
lacI(amber1)xF’
lacI(amber2)
How frequent are LacI+ relative to
distance separating the two mutations?
Would expect that mutations that are
close together would recombine
less that ones that are far apart
![Page 22: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/22.jpg)
Location of mutations in lacI that give different lacI phenotypes
![Page 23: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/23.jpg)
cAMP levels during growth of E. coli
![Page 24: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/24.jpg)
Effect of cAMP on LacZ induction
Note that cAMP has less effect on induction on poor carbon sources than on glucose—Why is this?
![Page 25: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/25.jpg)
cAMP alleviates
diauxic growth
![Page 26: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/26.jpg)
Adenyl cyclase activities of various E. coli strains
![Page 27: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/27.jpg)
![Page 28: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/28.jpg)
Effect of cAMP on cya mutants
1100110053365336
![Page 29: Standard recombination between an F’ and the chromosome.](https://reader035.fdocuments.net/reader035/viewer/2022070413/5697bfee1a28abf838cb98da/html5/thumbnails/29.jpg)
Correlation of physical and genetic maps
Answers “where are mutations located in a particular piece of genetic material”