Part:BBa_K4162023

ribozyme + T7_RBS + ybbO

Introduction

The ybbO gene under hammerhead ribozyme, with T7 RBS Part:BBa_K4162006 between them. This part can be used to assemble polycistrons in Escherichia coli.

Characterization

SDS-PAGE

Figure 1. SDS-PAGE. IPTG(-/+) = without/with 0.2 mM IPTG for 3-6 hours, adding IPTG to a bacteria culture with OD600 0.2-0.3. M: Protein molecular weight marker ladder. Lane 2~5、7～10: pET28 plasmids encoding ybbO，BCMO without any tag were transformed into DH5α strain, single clones (ybbO1, ybbO2, BCMO1, BCMO2) were picked for liquid LB culture. Protein expression was induced in parallel cultures by IPTG. Bacterial cultures were monitored by OD600, and 5x10^7 cells were harvested by centrifugation and lysis in 1x SDS sample buffer. Equal amount (10 μL, 2x10^6 cells) of whole cell lysate were analysed by SDS-PAGE (14% separation gel). Purple arrows point to ybbO protein.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 109
  Illegal AgeI site found at 702
  
• 1000
  COMPATIBLE WITH RFC[1000]