Reporter
GlpD

Part:BBa_K4140014

Designed by: mahmoud sayed   Group: iGEM22_AFCM-Egypt   (2022-10-01)
Revision as of 19:04, 11 October 2022 by Mohamed-Ab0elghare (Talk | contribs) (Usage)

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glpD

Part Description

Glycerol-3-phosphate dehydrogenase is a novel reporter gene for E. coli, the glpD gene outperforms well-known reporter genes like lacZ and gusA. Also, it has been investigated whether the glpD gene, which codes for glycerol-3-phosphate dehydrogenase, could function as an original E. coli reporter gene. In particular for transcripts with low translational efficiency, it could be demonstrated that the glpD transcript has significantly higher stability than the lacZ transcript. It might also be demonstrated that, unlike GusA, GlpD can be selected for in vivo activity through growth on glycerol.

Usage

We intended to employ this novel reporter gene to analyse the activity of the promoter of our therapeutic circuit in vitro as this reporter gene shows remarkable results by comparing it to the other reporter genes such as LacZ.

Literature Characterization

The two reporter genes gusA and glpD, which have better translational capacity than guaB, produced similar outcomes. The potential to select GlpD's in vivo behavior in synthetic media with glycerol as the sole carbon source is the main distinction between GusA and GlpD. This benefit of the glpD reporter gene has been effectively used to select non-canonical 5′-UTRs with high translational efficiency out of a random pool.

Figure.1 analysis of glpD activity by comparing it to LacZ and gusA
Figure 2. This table demonstrates the transcript level of the three reporter genes being compared






















Refrences

1.The art of reporter proteins in science: past, present and future applications BMB Rep., 43 (2010), pp. 451-460

2.The stability of Escherichia coli lacZ mRNA depends upon the simultaneity of its synthesis and translation EMBO J., 14 (1995), pp. 3252-3261

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 742
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 742
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 12
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 742
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 742
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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