Part:BBa_K4162031

ribozyme + T7_RBS + BCMO-StayGold

Introduction

Coding sequence of BCMO tagged with StayGold under hammerhead ribozyme, with T7 RBS Part:BBa_K4162006 between them. This part can be used to assemble polycistrons in E. coli.

We use this biobrick to verify the expression BCMO, which could not be induced and seen by SDS-PAGE. Please check our measurement page for details.

Figure 1. Fluorescent imaging revealed that IPTG induction increases the expression of both BCMO and ybbO. We observed fluorescence images of Hi-control Rosetta strain (1. Bacteria expressing BCMO-linker-StayGold; 2. Bacteria expressing BCMO and ybbO-tdMCP-GFP) through an Olympus fluorescence microscope. Samples induced with IPTG have stronger signal than controls.

Figure 2. Fluorescent imaging confirm the expression of both BCMO and ybbO. We observed fluorescence images of E. coli (1. Bacteria expressing BCMO-linker-StayGold ; 2. Bacteria expressing BCMO and ybbO-tdMCP-GFP ) through an Olympus fluorescence microscope with a 1.45 NA 150× oil objective. GFP and StayGold were excited by a 488 nm laser. Images were taken by the same settings for comparison.

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 957
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1601
  Illegal BsaI.rc site found at 1621