Difference between revisions of "Part:BBa K1592000"
ShuyanTang (Talk | contribs) |
ShuyanTang (Talk | contribs) |
||
| Line 20: | Line 20: | ||
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. | 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|600px|thumb|center|Php4d+LIP2 prepro-Mcfp+terminator]] | + | [[File:HUST-China_2015_circuit_1.png|600px|thumb|center|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. | 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|600px|thumb|center|Figure1: Protein Electrophoresis of LIP2-Mcfp3 (control: the train without plasmid). Mcfp3 protein is about 12kDa.]] | + | [[File:HUST-China_2015_results_4.jpg|600px|thumb|center|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. | 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. | ||
Revision as of 06:51, 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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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.
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.
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.