NYMU Taipei/Materials
From 2007.igem.org
Contents |
What we need in our project
Beginning of Story
- De-Ming Lin had concluded what a insulin-secretory cell should be from interature review
- Cells engineered for insulin expression should contain
- Proteases for insuline or proinsulin or express two chains (A, B) separately (activate insulin)
- glucose-sensing system (sense glucose)
- secretory granules that can release insulin promptly by exocytosis in response to extracellular glucose levels
Action plan
- Chih-Hsien Yang had concluded again in meeting record 2007-0716
- sensing glucose
- glucose-responsive promoter
- two component system for glucose sensing in E.coli
- iGEM 2007 parts and basic system configuration
- OmpR binding site
- Part:BBa_R0082 (15P, plate 1)
- Part:BBa_R0083 (17H, plate 1)
- Part:BBa_R0084 (11H, plate 1)
- EnvZ and RcsC fusion construct
- OmpR binding site
- lac operon in E.coli
- native insulin promoter in beta cell of eukaryotic system
- native beta cell can response glucose to secret insulin
- Regulation of insulin gene expression and insulin production in Nile tilapia (Oreochromis niloticus), 2007 General and Comparative Endocrinology
- glucose-responsive promoter
- producing insulin
- insulin included plasmid from
- domestic sources
- g1005043C07: MGC Human Verified Full-length cDNA Collection (IRAU), IMAGE id=3950204 (from BCRC, 4,000 NTD)
- Construct details
- 生科系蔡亭芬老師 可提供老鼠 (BL_4) 胰臟組織做 RT-PCR 取 insulin clone (連絡日期 8/3, PM 4:28)
- 8/6 蔡老師博士班學生吳家瑜 (分機 5529) 會幫忙抽好 RNA, 需要訂購 primer
- 8/7 拿到RNA, 已存入 -80 度冰箱
- 生理所阮琪昌老師: they don't have
- Library Resources Services at Gene Expression Analysis Center, VYMGC
- 訂購流程 and find genes by MGC finder
- 8/14 送出, 8/17 預定送來
- foreighn sources
- DNA synthesis or gene cloning
- iGEM registry search (iGEM part registry and iGEM 2007 kit)
- insulin antibody
- domestic sources
- insulin included plasmid from
- secreting insulin
- insulin transportation
- by signal peptide for protein sorting
- by vesicle for material transport in eukaryotic cell
- insulin transportation
- activating insulin
- Proteases (convertase PC2 and PC1/3) for insuline or proinsulin or
- express two chains (A, B) separately (without chain C)
- regulating insulin
- To balance the overexpressed insulin under low glucose supplement
- insulin protein inhibitor IGFBP7: Inhibition of Insulin Receptor Activation by Insulin-like Growth Factor Binding Proteins, 1997 JBC (posted by 迪璿)
- inhibitory anti-sense RNA for insulin mRNA (posted by Chih-Hsien)
- [Comparison of insulin regulatory models]
- To balance the overexpressed insulin under low glucose supplement
- diabetic mammlian cell line
- domestic sources
- foreighn sources
RNA Switch Biosensor
- Influenza virus like particle (VLP)-get from professor Ho, Mei-Shang (Academia Sinica, IBMS )
- iNOS promoter and iNOS gene construct-get from professor Lin, Wan-Wan (NTU)
- pTREX -GFP construct-get from professor Stephen Elledge (Harvard University)
- Cytochrome c- from VYMGC
- Antiswitch, TetR, pTet-get from iGEM
Make for practical use
- control the population of E.coli-based insulin secretory cell
- prevent the immune response