Difference between revisions of "Part:BBa K3452000"
Howie Chow (Talk | contribs) (→Contribution) |
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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> | ||
| − | Summary: We have successfully expressed hMT-1A in E. coli DH5α(Figure1 a). The OD600 of E.coli | + | 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> |
<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> | <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> | ||
<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> | <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> | ||
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）
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 946
Illegal SpeI site found at 1135
Illegal PstI site found at 1155 - 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 946
Illegal SpeI site found at 1135
Illegal PstI site found at 1155 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 862
Illegal BglII site found at 1051
Illegal BamHI site found at 673 - 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 946
Illegal SpeI site found at 1135
Illegal PstI site found at 1155 - 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]