Difference between revisions of "Part:BBa K1592000"
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<partinfo>BBa_K1592000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1592000 SequenceAndFeatures</partinfo>
  
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[[File:HUST_result4.jpg|thumb|450px|link=|frameless|center|'''Fig.3-1''' P<sub>cons</sub>+RBS+PBAN''(Mamestra brassicae'')]]
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[[File:HUST_part2.]]
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<h1>'''Application of the part'''</h1>
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This signal tag, LIP2 prepro, fused to Mcfp3 and constructed a new biobrick (BBa_K1592003), can lead the co-translational translocation of heterologous protein Mcfp-3 to be secreted out of the cell to achieve its function.
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[[File:HUST-China_2015_circuit_1|600px|thumb|center|Php4d+LIP2 prepro-Mcfp+terminator]]
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We concentrated cell culture solution of test Y.lipolytca JMY1212 cells transformed LIP2 prepro-Mcfp3 plasmid and control wildtype cells, and then separated proteins by SDS-PAGE.
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[[File:HUST-China_2015_results_4|600px|thumb|center|Figure1: Protein Electrophoresis of LIP2-Mcfp3 (control: the train without plasmid). Mcfp3 protein is about 12kDa.]]
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Figure shows an obvious ~12kDa protein bands of LIP2 prepro-Mcfp3 in test lane, which cannot be found in control lane. This result proves that LIP2 prepro signal peptide can successfully lead the flocculating proteins Mcfp3 into surrounding environment.
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===Functional Parameters===
 
===Functional Parameters===
 
<partinfo>BBa_K1592000 parameters</partinfo>
 
<partinfo>BBa_K1592000 parameters</partinfo>
 
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Revision as of 06:49, 14 September 2015

LIP2 prepro(signal peptide)


13 aa pre/4 XA or XP dipeptides/10 aa pro/KR cleavage site The pre-region of lipase 2 from Yarrowia lipolytica corresponds to the signal sequence. LIP2 prepro consist of 13aa pre-region of lipase2, four XA or XP dipeptides, 10aa pro-region of lipase2, and KR cleavage site. the dipeptides XA and XP are substrates for diamino-peptidase; the dibasic KR cleavage site is substrate for Xpr6p endoproteinase.

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]


Application of the part

This signal tag, LIP2 prepro, fused to Mcfp3 and constructed a new biobrick (BBa_K1592003), can lead the co-translational translocation of heterologous protein Mcfp-3 to be secreted out of the cell to achieve its function.

File:HUST-China 2015 circuit 1
Php4d+LIP2 prepro-Mcfp+terminator

We concentrated cell culture solution of test Y.lipolytca JMY1212 cells transformed LIP2 prepro-Mcfp3 plasmid and control wildtype cells, and then separated proteins by SDS-PAGE.

File:HUST-China 2015 results 4
Figure1: Protein Electrophoresis of LIP2-Mcfp3 (control: the train without plasmid). Mcfp3 protein is about 12kDa.

Figure shows an obvious ~12kDa protein bands of LIP2 prepro-Mcfp3 in test lane, which cannot be found in control lane. This result proves that LIP2 prepro signal peptide can successfully lead the flocculating proteins Mcfp3 into surrounding environment.