Composite

Part:BBa_K5115036

Designed by: Liyue Chen   Group: iGEM24_Fudan   (2024-09-19)
Revision as of 03:54, 1 October 2024 by Chenliyue (Talk | contribs)


ribozyme+RBS+Hpn+stem-loop

contributed by Fudan iGEM 2023

Introduction

This composite part is composed of hpn and improved ribozyme-assisted polycistronic co-expression system: pRAP. The hpn is a protein which can bind with nickel ions, get details in BBa_K1151001(Histidine-rich metal-binding protein)

Usage and Biology

The ribozyme-assisted polycistronic co-expression system can ensure that each cistron can initiate translation with comparable efficiency. For more information, please check part wiki of 2022 Fudan iGEM.

Characterization

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contributed by Fudan iGEM 2024
Figure 1: Comparison of E. coli Growth curve with and without hpn in 20 mg/L Ni²⁺

The graph illustrates the effect of Ni²⁺ on the growth of E. coli expressing hpn compared to E. coli without hpn expression in a medium containing 20 mg/L Ni²⁺ (E.coli strain: BL21 DE3, induced with 1 mM IPTG). The optical density (OD₆₀₀) of the initial bacterial suspension was adjusted to 0.5, and equal volumes of the suspension were added to each tube. E. coli growth was measured by OD₆₀₀, and the bacterial counts were calculated using a standard conversion, where OD₆₀₀ = 1 corresponds to 5.39 × 10⁸ cells. The results indicate that E. coli expressing Hpn has greater tolerance to Ni²⁺, exhibiting higher growth rates than E. coli without hpn expression under the same conditions.

contributed by Fudan iGEM 2024
Figure 2: Comparison of E. coli Growth curve with and without hpn in 50 mg/L Ni²⁺

The graph illustrates the effect of Ni²⁺ on the growth of E. coli expressing hpn compared to E. coli without hpn expression in a medium containing 50 mg/L Ni²⁺ (E.coli strain: BL21 DE3, induced with 1 mM IPTG). The optical density (OD₆₀₀) of the initial bacterial suspension was adjusted to 0.5, and equal volumes of the suspension were added to each tube. E. coli growth was measured by OD₆₀₀, and the bacterial counts were calculated using a standard conversion, where OD₆₀₀ = 1 corresponds to 5.39 × 10⁸ cells. The results indicate that E. coli expressing Hpn has greater tolerance to Ni²⁺, exhibiting higher growth rates than E. coli without hpn expression under the same conditions.

contributed by Fudan iGEM 2024
Figure 3: Comparison of E. coli Growth curve with and without hpn in 100 mg/L Ni²⁺

The graph illustrates the effect of Ni²⁺ on the growth of E. coli expressing hpn compared to E. coli without hpn expression in a medium containing 100 mg/L Ni²⁺ (E.coli strain: BL21 DE3, induced with 1 mM IPTG). The optical density (OD₆₀₀) of the initial bacterial suspension was adjusted to 0.5, and equal volumes of the suspension were added to each tube. E. coli growth was measured by OD₆₀₀, and the bacterial counts were calculated using a standard conversion, where OD₆₀₀ = 1 corresponds to 5.39 × 10⁸ cells. The results indicate that E. coli expressing hpn has greater tolerance to Ni²⁺, exhibiting higher growth rates than E. coli without hpn expression under the same conditions.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

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Categories
Parameters
None