Part:BBa_K5299005

RK600 helper plasmid

Introduction

pRK600 carries the tra genes, which are necessary for conjugative transfer. These genes enable the mobilization of plasmids between bacteria, facilitating the transfer of the transposon-bearing plasmid from the donor strain to the recipient strain via conjugation [1].

Usage and Biology

The RK600 plasmid, used in E. coli HB101, plays a key role in the mini Tn-7 system by carrying the tra genes necessary for conjugative transfer. These genes enable the mobilization of plasmids between bacteria, facilitating the transfer of the transposon-bearing Tn7 plasmid from a donor strain, like E. coli PIR2, to a recipient strain, such as Pseudomonas putida KT2440. In this system, pRK600 acts as a "helper" plasmid, providing the essential machinery for efficient conjugation, ensuring that the Tn7-M plasmid is successfully transferred, a process that would otherwise be inefficient without the involvement of pRK600 [2].

Figure 1: pRK600 in E. coli HB101 plate, iGEM Thessaly 2024.

Source

Dr. Esteban Martinez, a postdoctoral researcher at Victor de Lorenzo's Lab, provided us with the E.coli HB101 strain with the RK600 plasmid

References

[1] Zobel, S., Benedetti, I., Eisenbach, L., de Lorenzo, V., Wierckx, N., & Blank, L. M. (2015). Tn7-Based Device for Calibrated Heterologous Gene Expression in Pseudomonas putida. In ACS Synthetic Biology (Vol. 4, Issue 12, pp. 1341–1351). American Chemical Society (ACS). https://doi.org/10.1021/acssynbio.5b00058

[2] Salgado, S., Hernández-Herreros, N., & Prieto, M. A. (2024). Controlling the expression of heterologous genes in Bdellovibrio bacteriovorus using synthetic biology strategies. Microbial biotechnology, 17(6), e14517. https://doi.org/10.1111/1751-7915.14517

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]