Composite

Part:BBa_K2967013

Designed by: Shuang Wu   Group: iGEM19_NEU_CHINA   (2019-09-30)
Revision as of 15:34, 8 October 2019 by Hatsuse (Talk | contribs)


Amplifier system with hIL-10.

Amplifier system is created by our team NEU-CHINA 2019. To make sure that amplifier is able to strengthen the expression of gene downstream to promoter hrpL. A coomassie brilliant blue is demonstrated to show that more protein at 35kDA, which should be IL-10, is witnessed when introducing IL-10 gene downstream of promoter hrpL. And it can be seen that lanes with amplifier seems to be darker, which means that this amplifier system tends to strengthen overall protein expression.

T--NEU_China--part--amp_passway_use_this.png

Figure 1. Diagram of expressing vector. HrpR and HrpS amplifier system is used as a booster to boost the expression level of gene downstream promoter hrpL. Protein HrpR and HrpS can bind together to induce the PhrpL which strengthen the expression of downstream gene YebF-IL10.

800px-T--NEU_China--part--amp%2Bil-10-1.png

Figure 2. Expressing vector is transformed into BL21 strain. CFU is then inoculated to LB medium followed by 12h incubation at 37℃. After induction of IPTG, final concentration at 1mM, the culture is incubated at 37℃ for 2h. A coomassie brilliant blue is demonstrated to show the result. The red box shows where the target protein is.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2082
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1292
    Illegal SapI.rc site found at 1925


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