Composite

Part:BBa_K4183003

Designed by: Junjie Zhu   Group: iGEM22_HS_China   (2022-09-30)
Revision as of 14:39, 11 October 2022 by WSA-JACK (Talk | contribs) (→‎Usage and Biology)


Used for comparison with BBa_K4183002 to test the efficiency of secreted signal peptide.

Used for comparison with BBa_K4183002 to test the efficiency after the addition of the secreted signal peptide at the N-terminal end of lacZ.

Usage and Biology

We coated the engineered EcN bacteria containing BBa_K4183003 on LB agar plates containing IPTG as well as X-Gal and incubated them overnight at 37°C. The results showed that the breakdown of X-Gal could be clearly observed around the monoclonal colonies (Figure 1), which proved that lacZ was secreted extracellularly.

Figure1. Micrographs of X-Gal disassembled on LB agar plates containing X-Gal and IPTG. (A) When the N-terminal end of lacZ did not contain kp-sp, no disassembly of X-Gal was clearly observed around the monoclonal colonies. (B) When the N-terminal end of lacZ contains kp-sp, X-Gal is clearly observed to be disassembled around the monoclonal colony, showing a blue color.

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]


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Categories
Parameters
None