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