Part:BBa_K3452000

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）

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]