Difference between revisions of "Part:BBa K3452000"
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Group: iGEM20_CSU_CHINA (2020-10-26)<br>
 
Group: iGEM20_CSU_CHINA (2020-10-26)<br>
 
Author: Hao Zhou<br>
 
Author: Hao Zhou<br>
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Summary: We used the SmtA protein constructed by Groningen's team in 2009 to adsorb cadmium ions from outside the cell. We have successfully expressed protein in E. coli DH5α and supplemented experimental data to demonstrate that SmtA can Improve cadmium tolerance of E. coli respectively.（Figure a、b）<br>
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Summary: We have successfully expressed hMT-1A in E. coli DH5α(Figure1 a). Supplemented experimental data demonstrate that hMT-1A can improve cadmium tolerance of E. coli respectively and indirectly improve the ability to absorb cadmium.（Figure1. b, d, e）<br>
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<center>https://2020.igem.org/wiki/images/a/ae/T--CSU_CHINA--figure-engineering_success.png</center><br>
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<center>https://2020.igem.org/wiki/images/thumb/a/ae/T--CSU_CHINA--figure-engineering_success.png/800px-T--CSU_CHINA--figure-engineering_success.png</center><br>
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<center>Figure a：Western blotting results indicating the successful expression in E.Coli<br>
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<center>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-expressed E. coli, (e) Growth curves of MntH or MntH&TMCd1&APX2-expressed E. coli, (f) Cadmium uptake curve of MntH&TMCd1&APX2-expressed E. coli.</center><br>
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Figure b：Growth curve after successful protein expression</center><br>
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 18:20, 27 October 2020


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). Supplemented experimental data demonstrate that hMT-1A can improve cadmium tolerance of E. coli respectively and indirectly improve the ability to absorb cadmium.（Figure1. b, d, 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-expressed E. coli, (e) Growth curves of MntH or MntH&TMCd1&APX2-expressed E. coli, (f) Cadmium uptake curve of 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]