Regulatory

Part:BBa_K3020001

Designed by: WANG WENJIA   Group: iGEM19_BIT   (2019-09-06)
Revision as of 14:33, 6 September 2019 by WANGWENJIA (Talk | contribs)


recA promoter-Respond to sos response and initiate expression of downstream repair proteins

The SOS response is an inducing response when DNA replication is blocked or damaged. In E. coli, this reaction is regulated by the recA-lexA system, which does not function under normal physiological conditions. When DNA replication is blocked or damaged to produce an exposed single strand, the protease function of one of the RecA functions is immediately activated. The repressor protein LexA spontaneously degraded and sheds from the promoter of the SOS gene, thereby promoting the expression of genes involved in the SOS response (such as uvrA, uvrB, uvrC, uvrD, ssb, recA, and recN). Thereby, the functions of excision repair, post-replication repair and strand break repair related to these genes are generated, and a series of gene level and cell level responses are exerted.The expression levels of these SOS genes range from several times to tens of times when the SOS reaction does not occur. After the elimination of the inducing factor (such as a large number of DNA single strands), the protease activity of RecA disappears, and the amount of LexA protein is significantly increased, and the repression is re-acted. When the SOS reaction occurs, it can cause an increase in damage repair function in a short period of time. Since the expression of the SOS gene is closely related to the viability of its promoter, the signal of the reporter gene constructed under the gene promoter can indicate the activity of the promoter, and the signal size of the reporter factor and the concentration of the DNA damage reagent usually have a certain dose-effect relationship. Therefore, the SOS Promoter + Reporter System can be used as an indicator of the ability of a compound to detect DNA damage.This sequence is the promoter and RBS base sequence of the recA repair protein.

The recA promoter does not have a strict sequence range on the NCBI website. Therefore, when using the PCR to clone the target fragment, we designed the primer and completely cloned the recA repair protein base ATG upstream of 390 bp.

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]


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Categories
Parameters
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