Upload
ross
View
32
Download
0
Embed Size (px)
DESCRIPTION
Summer Research Inhibition of Type I Interferon Signal Transduction Pathways by JEV infection. 中研院生醫所 R443 林宜玲老師 清大生科 05 蔡文揚 2003. 實驗室簡介. 研究 Flavivirus- 日本腦炎病毒 登革熱病毒 黃質病毒的基因體為單股正向的 RNA ,能夠作出一個大蛋白質,經過切割後形成大約 11 個病毒結構性以及非結構性蛋白質。. *. *. C. E. NS1. NS3. NS5. NS2A. NS2B. NS4A. - PowerPoint PPT Presentation
Citation preview
Summer Research Inhibition of Type I Interferon Signal Transduction Pathways by
JEV infection
中研院生醫所 R443林宜玲老師
清大生科 05 蔡文揚 2003
實驗室簡介• 研究 Flavivirus- 日本腦炎病毒 登革熱病毒• 黃質病毒的基因體為單股正向的 RNA ,能夠作出一個
大蛋白質,經過切割後形成大約 11 個病毒結構性以及非結構性蛋白質。
CprM
/M E NS1NS1
NS2ANS2B
NS3NS4A
NS4BNS5
: host signalase: viral protease NS2B-3: furin or other golgi-localized protease: unknown protease
* *
Interferons (IFNs) are a family of cytokines that possess antiviral activity.
Type I IFNs (viral IFNs)
Type II IFN (immune IFN
include IFN-, IFN-, IFN- and IFN-
produced by most types of cell can produce
IFN-
produced only by NK cells, CD4+ T help cells and CD8+ cytotoxicity cells
IFNs-mediated signaling pathway and anti-viral strategies
Viral interference with the IFN system
Our works
• Construct JEdomainE/pIRES-hrGFP
• Construct prMM/ME/pR21-TE12Q
Construct JEIII/pIRES-hrGFP
pSecTagA pSecTagApCR3.1JEV-E IIIpCR3.1
BamH1 Xho1Kpn1
Asp7181
Hind IIISfi1
Nhe1
Xba1
Apa1
Sal1
ATG Myc epitope
pIRES hrGFP
Sma1 Sal1
JEV-E III/pSecTagA
Nhe1
T4 DNA polymerase
Clean up
Sal1
IRES-hrGFP
Sma1(blunt end)
Clean up
Sal1
Clean up
CIP
Ligation
Ligase
Transformation
Gel elution
Transfection
• Lipofectamine (artificial membrane)
• BHK-21
Immunofluorescence Assay
Western plot
IFAIRES-hrGFP control
JE-III/pIRES-hrGFP
DAPI
DAPI
Myc+Texas Red
Myc+Texas Red
Construct prMM/ME/pr21-TE12Q
• Fail………….
心得• 科學家一定要心地善良