Part:BBa_K5115036

ribozyme+RBS+Hpn+stem-loop

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

Growth curve of E.coli

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.

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.

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]