Part:BBa_K629001

recAp, could be started with exposure to irradiation, UV, nalidixic acid

Background

The recA gene of Escherichia coli is involved in genetic recombination, in postreplication repair, and in a number of other cellular functions such as mutagenesis, phage induction, and cell division. RecA protein is related to SOS system-- DNA repair, which is activated by recA promoter. The active level of recA promoter is fluctuates from low basal levels to very high levels after treatments that damage DNA or stall DNA replication. In our project, recA promoter is used to detect single-chain DNA which result from ionizing radiation, x-ray, ultraviolet radiation and some kinds of drug. According to the charater, ionizing radiation can activate recA promoter and raise the level of gene expression which under recA promoter's regulation.

Two exchangable ways to start recAp

Because of the special pathway in start of recAp, which is small pieces of break DNA can lead to the SOS system, there has been several ways to start this promoter from research and our try. In this case, the range of usage of this promoter could be spread.

1. UV light This way is based on the basic theroy of start of recAp, which means that when UV light damage DNA into some small pieces, recAp would be started.

What a pity in this method is that the parameters cannot be determined for the unstable data.

2. Nalidixic Acid

Nalidixic acid selectively and reversibly blocks DNA replication in susceptible bacteria. Nalidixic acid and related antibiotics inhibit a subunit of DNA gyrase and induce formation of relaxation complex analogue. It also inhibits the nicking dosing activity on the subunit of DNA gyrase that releases the positive binding stress on the supercoiled DNA. Nalidixic acid is effective against both gram-positive and gram-negative bacteria. In lower concentrations, it acts in a bacteriostatic manner; that is, it inhibits growth and reproduction. In higher concentrations, it is bactericidal, meaning that it kills bacteria instead of merely inhibiting their growth.

All in all, nalidixic acid inhibits DNA gyrase, blocking DNA replication and leading to degradation of DNA. As a result, it could be an optimal alternative for nuclear radiation. As a result, it has ability to start recAp.

Reference

[1] S. Nuyts et al., Radiation Research. 155, 716 (2001). [2] S. I.Feinsteinl et al., Nucleic Acids Research. 11, 2927 (1983).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]