Difference between revisions of "Part:BBa K3452000"

Revision as of 20:52, 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 expressed by HMT-1A represents that the concentration of bacteria, 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. c). , 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 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>
+Summary:  We have successfully expressed hMT-1A in E. coli DH5α(Figure1 a). The OD600 of E.coli expressed by HMT-1A represents that the concentration of bacteria, 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. c). , 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>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>