Coding

Part:BBa_K4451010

Designed by: Brooks J Rady   Group: iGEM22_Sheffield   (2022-09-30)


T4 AsiA

BBa_K4451010 encodes the AsiA protein from the T4 phage. This protein has been reported to interact with sigma-70 factor and inhibits recognition of sigma-70 promoters (Ouhammouch et al., 1995).

Description

A transcriptional inhibitor from T4 phage, which causes growth rate suppression in E. coli. The AsiA gene encodes an anti-σ factor, which is able to bind to σ70 and prevent the RNA polymerase σ70 holoenzyme from recognising σ70 specific promoters, thus enacting a transcriptional downregulation of housekeeping genes and subsequently causing growth rate suppression. AsiA further enables the transcription of phage middle genes as part of the T4 infection cycle (J. Liu et al., 2004).

bba-k4451010-meanlog2od.png


Figure 1: Mean log2(OD) against time for AsiA, concentrations refer to the [IPTG] used to induce the pLac promoter regulating AsiA.


Figure 1 shows that there's a significant decrease in growth of E. coli when they are transformed with a plasmid containing AsiA. The decreased mean log 2 OD observed without induction of pLac shows leaky expression of the growth slower gene has an impact on bacterial cell growth. The greater the [IPTG], the larger the decrease in growth rate in the case of 4uM and 1000uM AsiA, this is not observed for concentration changes between 2 and 4uM, suggesting that in the future more [IPTG] need to be used between this interval 0f 4 and 1000uM. This supports that AsiA causes a growth slowing impact in E. coli, and the level of growth slowing can be controlled using an inducible promoter.

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
None