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Specific Inhibition of HSV-1 Capsid Protein and US12 Gene by RNA Interference in Vitro

Author: LiShen
Tutor: WangYiFei
School: Jinan University
Course: Microbial and Biochemical Pharmacy
Keywords: RNA interference Capsid protein QUICK US12 HSV-1
CLC: R373
Type: Master's thesis
Year: 2011
Downloads: 22
Quote: 0
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Abstract


Objective: To screen high - silence HSV-1 capsid gene siRNA ; study of siRNA silencing the target gene of HSV - I infection and proliferation , provide the basis for the anti- herpes simplex virus drug development ; explore the capsid protein and the host cell , the relationship between capsid protein signaling pathway ; screening and efficient silent HSV-1 US12 gene siRNA to study the impact of the proliferation of HSV - I the siRNA silencing US12 gene . Methods: design and chemical synthesis for seven HSV-1 capsid protein gene siRNA transfected Vero cells , the inhibitory effect of the target gene mRNA expression by quantitative PCR siRNA . The plaque reduction Experimental evaluation siRNA inhibition of target gene of HSV-1 infection and proliferation . Morphological observation of siRNA in infected cell disease . Transmission electron microscopy siRNA interference copy, and the impact of the nuclear viral capsid nuclear . The building can be used for the purpose of high-throughput screening of gene and protein expression of pEGFP-N1-US12 fusion expression plasmid . QUICK Research VP5 interacting protein . Results: cotransfection experiment screened efficiently inhibit the expression of UL18, UL19, UL26, UL26.5, UL35, UL38mRNA siRNA , but the plaque reduction was found only for the UL18 and UL19 genes of siRNA on the proliferation of HSV-1 was significantly inhibitory effect. siRNA interference UL19 gene could significantly inhibit cell lesions . Transmission electron microscopy showed interference UL19 gene can reduce the number of nuclear capsid , inhibit the release of the viral capsid . Successfully constructed fusion expression plasmid pEGFP-N1-US12 can be used for high-throughput screening of target gene and protein expression . QUICK technology to filter out the five protein VP5 interaction role . Conclusion: The design and synthesis of siRNA can effectively reduce the HSV-1 capsid protein gene expression , UL18 and UL19 siRNA can significantly inhibit viral infection and proliferation , suggesting that gene UL18 and UL19 gene is the development of anti- HSV-1 a potential target for antiviral drugs . RNA interference US12 gene can significantly reduce US12 mRNA level and fusion protein expression , but HSV - 1 , the in vitro proliferation of no significant impact , this indicates that the US12 expression Claim early protein ICP47 may HSV-1 in vitro proliferation are not directly related .

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CLC: > Medicine, health > Basic Medical > Medical Microbiology ( pathogenic bacteriology,pathogenic microbiology ) > Human Virology ( pathogenic virus)
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