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). 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）<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 / hMT-1A / MntH & TMCd1 & APX2)-expressed E. coli, (e) Growth curves of ( MntH / hMT-1A / 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

Latest revision as of 21:05, 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). 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]
    Unknown
  • 12
    INCOMPATIBLE WITH RFC[12]
    Unknown
  • 21
    INCOMPATIBLE WITH RFC[21]
    Unknown
  • 23
    INCOMPATIBLE WITH RFC[23]
    Unknown
  • 25
    INCOMPATIBLE WITH RFC[25]
    Unknown
  • 1000
    COMPATIBLE WITH RFC[1000]