Part:BBa_K1906005

Beta-subunit of Qbeta replicase holoenzyme

This protein is the beta subunit of Qbeta replicase.

Characterlization

It was quite surprising that plasmids contain this CDS part don't lead to a heavy production of the protein upon the induction. However, this situation can be improved when EF-Ts and EF-Tu are co-expressed with beta subunit.

Figure 1 is the SDS-PAGE result comparison between cells that were transformed with these three proteins and beta subunit alone.

Figure 1. SDS-PAGE result comparison between three-protein-expressing cells and beta subunit-expressing cells. Wild-type cells were included as a control group. The gel on the left shows the results of cells that had been induced with 2mM IPTG and the gel on the right shows the results of same groups of cells that had not been induced. The blue arrow indicates the band position of beta subunit.

It can be clearly seen that cells transformed with beta subunit alone had no detectable level of protein expression. However, the expression of beta subunit can be greatly restored when plasmids that carry EF-Tu and EF-Ts are transformed into the cells.

Usage and Biology

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1317
  Illegal BamHI site found at 1460
  Illegal XhoI site found at 991
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1473
  
• 1000
  COMPATIBLE WITH RFC[1000]