Part:BBa_K5210011

RecO

In Deinococcusradiodurans, RecF and RecR are required to stabilize and maintain replication forks arrested by UV-induced DNA damage. RecO functions with RecF and RecR to protect the nascent lagging strand of arrested replication forks after UV-irradiation[1]. In the absence of RecO, the nascent DNA at arrested replication forks is extensively degraded and replication fails to recover.

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

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 381
  Illegal BamHI site found at 477
  Illegal BamHI site found at 492
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]