Part:BBa_K5210012
RecF
In Deinococcusradiodurans, RecF and RecR are required to stabilize and maintain replication forks arrested by UV-induced DNA damage[1]. In the absence of RecF, replication fails to recover, and the nascent lagging strand of the arrested replication fork is extensively degraded by the RecQ helicase and RecJ nuclease. recO mutants are epistatic with recF and recR with respect to recombination and survival assays after DNA damage.
Usage and Biology
RecO and RecF of Deinococcus radiodurans have been applied to the construction of prokaryotic expression systems to confer radiation resistance to prokaryotes . In this way,we successfully constructed the microbial survival part.As for exploring the expression of RecO and RecF, we introduced the two genes into Pet-30b vector separately as two experimental groups, and induced them with 0.5 mM IPTG for 1h, 2h, 3h and 4h, respectively, and the same low-temperature induction was carried out to perform SDS-PAGE experiments, and the two groups were compared for observing the expression of the target proteins.
In addition, we verified the survival of E. coli introduced with RecO, F after different lengths of UV irradiation.
Experiment results
A. Plasmid construction
B. Changes in the survival of bacteria after UV irradiation at different doses
C. SDS-PAGE image
Reference
[1]Fernández, S et al. “Analysis of the Bacillus subtilis recO gene: RecO forms part of the RecFLOR function.” Molecular & general genetics : MGG vol. 261,3 (1999): 567-73. doi:10.1007/s004380051002 Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 226
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 226
Illegal NotI site found at 272 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 222
Illegal BamHI site found at 915 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 226
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 226
Illegal NgoMIV site found at 1024
Illegal AgeI site found at 736 - 1000COMPATIBLE WITH RFC[1000]
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