Difference between revisions of "Part:BBa K2922044"
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===Summary===
 
===Summary===
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This part is constructed for the cooperation of CAV1 and fusion protein DsbA-MBP. CAV1 and DsbA-MBP was connected in the same circuit to improve the absorption ability of E. coli toward lead in the aqueous environment through enrichment.
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This part was constructed for the cooperation of CAV1 and fusion protein DsbA-MBP. CAV1 and DsbA-MBP was connected in the same circuit to improve the absorption ability of ''E. coli'' toward lead in the aqueous environment through enrichment.
 
<table><tr><th>[[Image:Im-6.png|thumb|400px|Figure 1. DsbA fusion protein works together with CAV1]]</th><th></table>
 
<table><tr><th>[[Image:Im-6.png|thumb|400px|Figure 1. DsbA fusion protein works together with CAV1]]</th><th></table>
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===Characterization===
 
===Characterization===
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Similar to the description in BBa_K2922043, ICP-MS was used to accurately determine the amount of lead absorption per unit mass of cells. The amounts of lead absorbed by the bacteria with DsbA-MBP and DsbA-MBP-CAV1 were measured and compared.
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Similar to the description in <partinfo>BBa_K2922043</partinfo>, ICP-MS was used to accurately determine the amount of lead absorption per unit mass of cells. The amounts of lead absorbed by the bacteria with DsbA-MBP and DsbA-MBP-CAV1 were measured and compared.
 
The relative lead absorption capacity after incubation in medium with 50 μM Pb(II) was shown in Fig.2. The relative absorption capacity of DsbA-MBP-CAV1 increased significantly and was much higher than that of DsbA-MBP. At the same time, the death and rupture of bacteria caused by the toxicity of Pb(II) may be the reason for absorption capacity decreased at 18 h in DsbA-MBP group.
 
The relative lead absorption capacity after incubation in medium with 50 μM Pb(II) was shown in Fig.2. The relative absorption capacity of DsbA-MBP-CAV1 increased significantly and was much higher than that of DsbA-MBP. At the same time, the death and rupture of bacteria caused by the toxicity of Pb(II) may be the reason for absorption capacity decreased at 18 h in DsbA-MBP group.
 
<table><tr><th>[[Image:Im-7.png|thumb|300px|Figure 2. Relative Pb（II）Absorption Capacity Per Unit Mass（DsbA-MBP &DsbA- MBP-Cav1）]]</th><th></table>
 
<table><tr><th>[[Image:Im-7.png|thumb|300px|Figure 2. Relative Pb（II）Absorption Capacity Per Unit Mass（DsbA-MBP &DsbA- MBP-Cav1）]]</th><th></table>
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===Conclusion===
 
===Conclusion===

Latest revision as of 14:56, 21 October 2019


Co-expression system of BBa_K346030 and BBa_K2922042 for enhancing lead absorption

Summary

This part was constructed for the cooperation of CAV1 and fusion protein DsbA-MBP. CAV1 and DsbA-MBP was connected in the same circuit to improve the absorption ability of E. coli toward lead in the aqueous environment through enrichment.

Figure 1. DsbA fusion protein works together with CAV1

Characterization

Similar to the description in BBa_K2922043, ICP-MS was used to accurately determine the amount of lead absorption per unit mass of cells. The amounts of lead absorbed by the bacteria with DsbA-MBP and DsbA-MBP-CAV1 were measured and compared. The relative lead absorption capacity after incubation in medium with 50 μM Pb(II) was shown in Fig.2. The relative absorption capacity of DsbA-MBP-CAV1 increased significantly and was much higher than that of DsbA-MBP. At the same time, the death and rupture of bacteria caused by the toxicity of Pb(II) may be the reason for absorption capacity decreased at 18 h in DsbA-MBP group.

Figure 2. Relative Pb（II）Absorption Capacity Per Unit Mass（DsbA-MBP &DsbA- MBP-Cav1）

Conclusion

The results demonstrated that the introduction of CAV1 could not only enhance the lead absorption efficiency, but also improve the resistance of bacteria against heavy metal.


Reference

1. J. Shin et al., Display of membrane proteins on the heterologous caveolae carved by caveolin-1 in the Escherichia coli cytoplasm. Enzyme Microb Technol 79-80, 55-62 (2015).
2. J. Shin et al., Endocytosing Escherichia coli as a Whole-Cell Biocatalyst of Fatty Acids. ACS Synthetic Biology 8, 1055-1066 (2019).



Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 520
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 520
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 308
    Illegal XhoI site found at 1005
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 520
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 520
    Illegal AgeI site found at 140
  • 1000
    COMPATIBLE WITH RFC[1000]