Difference between revisions of "Team:HUST-China/Notebook"
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Revision as of 12:14, 17 September 2015
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Euk.cement
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Details
Standalization
Week 1 6.29-7.5
Week 2 7.6-7.12
Week 3 7.13-7.19
Week 4 7.20-7.26
Week 5 7.27-8.2
Week 6 8.3-8.9
PCR: More details, protocol WangJie Liu and JunJie Xu
Restriction Enzyme Digestion
WangJie Liu and JunJie Xu
DNA ligation&Transformation
WangJie Liu and JunJie Xu
Successfully align the sequencing results with the objective sequences:
Colony PCR
WangJie Liu and JunJie Xu
Extract the plasmid,Conserving.
Week 7 8.10-8.16
PCR:
More details, protocol
WangJie Liu
Restriction Enzyme Digestion
WangJie Liu
DNA ligation&Transformation
WangJie Liu
Colony PCR
Successfully align the sequencing results with the objective sequences:
WangJie Liu
Extract the plasmid,Conserving.
Week 9 8.24-8.30
Week 10 8.31-9.6
Successfully align the sequencing results with the objective sequences
Standalization of BD-CRY2 and AD-CIB1 finished.
Successfully align the sequencing results with the objective sequences
Standalization of CRY2 and CIB1 finished.
Light Control
Week 1 6.29-7.5
Week 2 7.6-7.12
Successfully align the sequencing results with the objective sequences of Pgal1 and CYC_TT.
Extract the plasmid.
Conserving.
Week 3 7.13-7.19
Week 4 7.20-7.26
Week 5 7.27-8.2
Successfully align the sequencing results with the objective sequences
Digestion Test
Guozhao Wu
Week 6 8.3-8.9
Successfully align the sequencing results with the objective sequences
Pgal-ROX1-CYC_TT circuit was constructed.
Week 7 8.10-8.16
Week 8 8.17-8.23
Week 9 8.24-8.30
Week 10 8.31-9.6
viscous parts
Week 1 6.29-7.5
Week 2 7.6-7.12
PCR:
More details, protocol
Wang Xi and JunJie Xu
Restriction Enzyme Digestion
Wang Xi and JunJie Xu
DNA ligation&Transformation
Wang Xi and JunJie Xu
Colony PCR
Successfully align the sequencing results with the objective sequences:
Wang Xi and JunJie Xu
Extract the plasmid,Conserving
Week 3 7.13-7.19
PCR:
More details, protocol
ZhangYu Cheng
Restriction Enzyme Digestion
ZhangYu Cheng
DNA ligation&Transformation
ZhangYu Cheng
Colony PCR
ZhangYu Cheng
Successfully align the sequencing results with the objective sequences:
Extract the plasmid,Conserving.
Week 4 7.20-7.26
PCR:
More details, protocol
Wang Xi and JunJie Xu
Restriction Enzyme Digestion
Wang Xi and JunJie Xu
DNA ligation&Transformation
Wang Xi and ZhangYu Cheng
Colony PCR
Wang Xi and JunJie Xu
Successfully align the sequencing results with the objective sequences:
Extract the plasmid,Conserving.
Tramsformation to yeast
Lei Yin
Genome extract and PCR test
Lei Yin
Positive strain conservation
Week 5 7.27-8.2
PCR:
More details, protocol
ZhangYu Cheng
Restriction Enzyme Digestion
ZhangYu Cheng
DNA ligation&Transformation
ZhangYu Cheng
Colony PCR
ZhangYu Cheng
Successfully align the sequencing results with the objective sequences:
Extract the plasmid,Conserving.
Trasnsform to yeast
Genome extract and PCR test
Lei Yin
Positive strain conservation
Week 6 8.3-8.9
PCR:
More details, protocol
ZhangYu Cheng
Restriction Enzyme Digestion
ZhangYu Cheng
DNA ligation&Transformation
ZhangYu Cheng
Colony PCR
ZhangYu Cheng
Successfully align the sequencing results with the objective sequences:
Extract the plasmid,Conserving.
Trasnsform to yeast
Lei Yin
Genome extract and PCR test
Lei Yin
Positive strain conservation
Week 7 8.10-8.16
SDS-PAGE
Lei Yin
Immunoflourecense
ZhangYu Cheng
Verification of the secreting Mcfp-3 (Coomassie blue staining)
ZhangYu Cheng
Week 8 8.17-8.23
Week 9 8.24-8.30
Week 10 8.31-9.6
Improvement
Week 1 6.29-7.5
Week 2 7.6-7.12
Week 3 7.13-7.19
Week 4 7.20-7.26
Week 5 7.27-8.2
Week 6 8.3-8.9
Week 7 8.10-8.16
Week 8 8.17-8.23
Week 9 8.24-8.30
Week 10 8.31-9.6
Consolidation
Week 1 6.29-7.5
Week 2 7.6-7.12
Week 3 7.13-7.19
Week 4 7.20-7.26
Week 5 7.27-8.2
Week 6 8.3-8.9
Week 7 8.10-8.16
Week 8 8.17-8.23
Week 9 8.24-8.30
Week 10 8.31-9.6
Host strain
We adopt a marine yeast Yarrowia lipolytca JMY1212 as our chassis for the specific application this year--building artificial reef. Our host is generally regarded as safe microorganism and has an abroad application prospect in industries of foods and medicine. Researches shows that Y.lipolytica is widely found in marine environment which exists stably in the ocean and healthy marine fishes’ intestine and it is able to make use of Glucose,ethanol,acetate or other hydrophobic substrate(alkanes,fatty acid and lipid ) as carbon source with fast growing capacity and adaptability for high-density fermentation. Moreover in the expression system of Y.lipolytica, transformed plasmids can integrate into multiple site and they are all genetically stable in the genome. And the signal peptide XPR2 pre,LIP2 prepro and so on are all demonstrated to own high efficiency of secreting protein.So it is obvious Y.lipolytica is a powerful host to express and secrete complex recombinant protein. Based on the above reasons,Yarrowia lipolytica are regarded as ideal strain for our marine application of micro-solidation
Improvement