Difference between revisions of "Part:BBa K2967013"
 
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<partinfo>BBa_K2967013 short</partinfo>
 
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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.
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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 was demonstrated that more protein at 35kDA, which should be IL-10, was witnessed when introducing IL-10 gene downstream of promoter hrpL. This amplifier system tended to strengthen overall protein expression including the human IL-10.
  
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https://2019.igem.org/wiki/images/d/df/T--NEU_China--part--amp_passway_use_this.png
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Figure 1. 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.
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'''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 strengthens the expression of downstream gene YebF-IL10.
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https://static.igem.org/mediawiki/parts/thumb/1/1b/T--NEU_China--part--amp%2Bil-10-1.png/800px-T--NEU_China--part--amp%2Bil-10-1.png
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'''Figure 2. Protein expression of human IL-10 gene which transformed in BL21 strain. CFU was then inoculated to LB medium followed by 12h incubation at 37℃.''' After induction of IPTG with the final concentration at 1mM, the culture was incubated at 37℃ for 2h. A coomassie brilliant blue was demonstrated to show the result. The red box shows where the target protein is.
  
 
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<!-- Add more about the biology of this part here

Latest revision as of 09:15, 16 October 2019


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 was demonstrated that more protein at 35kDA, which should be IL-10, was witnessed when introducing IL-10 gene downstream of promoter hrpL. This amplifier system tended to strengthen overall protein expression including the human IL-10.

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 strengthens the expression of downstream gene YebF-IL10.

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

Figure 2. Protein expression of human IL-10 gene which transformed in BL21 strain. CFU was then inoculated to LB medium followed by 12h incubation at 37℃. After induction of IPTG with the final concentration at 1mM, the culture was incubated at 37℃ for 2h. A coomassie brilliant blue was 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