Difference between revisions of "Part:BBa K3452000"

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）

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]

@@ Line 9: / Line 9: @@
 Group: iGEM20_CSU_CHINA (2020-10-26)<br>
 Author: Hao Zhou<br>
-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>
+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/a/ae/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>Figure a：Western blotting results indicating the successful expression in E.Coli<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>
-Figure b：Growth curve after successful protein expression</center><br>
 <!-- Add more about the biology of this part here