In Vitro Expression of Native H5 and N1 Genes of Avian Influenza Virus by Using Green Fluorescent Protein as Reporter
نویسندگان
چکیده
HARTAWAN, R., K. ROBINSON, T. MAHONY and J. MEERS. 2011. In vitro expression of native H5 and N1 genes of avian influenza virus by using Green Fluorescent Protein as reporter. JITV 16(3): 234-241. The hemagglutinin and neuraminidase are important immunogen of avian influenza virus that are suitable for recombinant experimentation. However, both genes have been experienced rapid mutation resulting in diverse variety of genotypes. Hence, gene expression in recombinant systems will be difficult to predict. The objective of the study was to examine expression level of H5 and N1 genes from a field isolate by cloning the genes into expression vector pEGFP-C1. Two clones respresenting fulllength of H5 and N1 gene in plasmid pEGFP-C1 were transfected into chicken embryo fibroblasts (CEF), rabbit kidney (RK13) and African green monkey kidney (VERO) cells using Lipofectamine ‘Plus’ reagent. The experiment showed level of gene expression in the VERO cell was higher than in the RK13 and CEF cells. Observations using fluorescent microscopy and Western blotting revealed that the N1 gene was expressed better in all cells compared to the H5 gene.
منابع مشابه
In vitro Labeling of Neural Stem Cells with Poly-L-Lysine Coated Super Paramagnetic Nanoparticles for Green Fluorescent Protein Transfection
Background: The magnetic nanoparticle-based transfection method is a relatively new technique for delivery of functional genes to target tissues. We aimed to evaluate the transfection efficiency of rat neural stem cell (NSC) using poly-L-lysine hydrobromide (PLL)-coated super paramagnetic iron oxide nanoparticles (SPION). Methods: The SPION was prepared and coated with PLL as transfection agent...
متن کاملMortality of wild swans associated with naturally infection with highly pathogenic H5N1 avian influenza virus in Iran
In the February 2006 in two wetlands in northern Iran, the mortality among wild swans was observed. Paralysis was the most prominent feature of the disease. Histologically, diffused necrosis of acinar cells in pancreas, degeneration and necrosis of some neurons in central nervous system (CNS), sever necrotic and hemorrhagic enteritis, foci of haemorrahge and myocardial cell necrosis in the he...
متن کاملA New Reporter Gene Technology: Opportunities and Perspectives
The paper summarizes the current status of the reporter gene technology and their basics. Reporter gene technology is widely used to monitor cellular events associated with gene expression and signal transduction. Based upon the splicing of transcriptional control elements to a variety of reporter genes, it “reports” the effects of a cascade of signaling events on gene expression inside cells. ...
متن کاملDevelopment and Evaluation of Real-Time Reverse Transcription Polymerase Chain Reaction Test for Quantitative and Qualitative Recognition of H5 Subtype of Avian Influenza Viruses
Avian influenza viruses (AIV) affect a wide range of birds and mammals, cause severe economic damage to the poultry industry, and pose a serious threat to humans. Highly pathogenic avian influenza viruses (HPAI) H5N1 were first identified in Southeast Asia in 1996 and spread to four continents over the following years. The viruses have caused high mortality in chickens and various bird species ...
متن کاملAn Enterovirus-Like RNA Construct for Colon Cancer Suicide Gene Therapy
Background: In gene therapy, the use of RNA molecules as therapeutic agents has shown advantages over plasmid DNA, including higher levels of safety. However, transient nature of RNA has been a major obstacle in application of RNA in gene therapy. Methods: Here, we used the internal ribosomal entry site of encephalomyocarditis virus and the 3’ non-translated region of Poliovirus to design an en...
متن کامل