Coding

Part:BBa_K3452000

Designed by: Hao Zhou   Group: iGEM20_CSU_CHINA   (2020-10-26)
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hMT-1A

Literature showed that the absorption capacity of recombinant strain to Cd increased to 7.90 μ mol/mg dry cell weight, which was nearly 19 times higher than that of the control strain. TMCd1 is made up of 156 amino acids, of which 47 are cysteine 2+ residues that can bind 16 Cd. Therefore, Suleman et al. expressed the TMCd1 gene fused to the glutathione S-transferase gene in E. coli BL21 DE3 cells. We used it in the cytoplasm to chelate the cadmium ions inhaled from the cytosol of E. coli.

Contribution

Group: iGEM20_CSU_CHINA (2020-10-26)
Author: Hao Zhou
Summary: We have successfully expressed hMT-1A in E. coli DH5α(Figure1 a). The OD600 of E.coli expressing hMT-1A represents that the concentration of E. coli, will increase faster than that without expression(Figure1. b). At the same time, it can maintain a high strain concentration at different cadmium concentrations(Figure1. d). , and indirectly shows a certain cadmium absorption ability.(Figure1. e)

800px-T--CSU_CHINA--figure-engineering_success.png

Figure1: (a) WB proves the successful expression of our protein, (b) Growth curves of chelated protein (SmtA, hMT-1A, TMCd1)-expressed E. coli, (c) Growth curves of APX2-expressed E. coli, (d) Cadmium uptake curve of (MntH / hMT-1A / MntH & TMCd1 & APX2)-expressed E. coli, (e) Growth curves of ( MntH / hMT-1A / MntH & TMCd1 & APX2)-expressed E. coli.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 946
    Illegal SpeI site found at 1135
    Illegal PstI site found at 1155
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 946
    Illegal SpeI site found at 1135
    Illegal PstI site found at 1155
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 862
    Illegal BglII site found at 1051
    Illegal BamHI site found at 673
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 946
    Illegal SpeI site found at 1135
    Illegal PstI site found at 1155
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 946
    Illegal SpeI site found at 1135
    Illegal PstI site found at 1155
    Illegal AgeI site found at 724
    Illegal AgeI site found at 892
  • 1000
    COMPATIBLE WITH RFC[1000]


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