title: Partial antiviral activities detection of chicken Mx jointing with neuraminidase gene (NA) against Newcastle disease virus.
abstract: As an attempt to increase the resistance to Newcastle Disease Virus (NDV) and so further reduction of its risk on the poultry industry. This work aimed to build the eukaryotic gene co-expression plasmid of neuraminidase (NA) gene and myxo-virus resistance (Mx) and detect the gene expression in transfected mouse fibroblasts (NIH-3T3) cells, it is most important to investigate the influence of the recombinant plasmid on the chicken embryonic fibroblasts (CEF) cells. cDNA fragment of NA and mutant Mx gene were derived from pcDNA3.0-NA and pcDNA3.0-Mx plasmid via PCR, respectively, then NA and Mx cDNA fragment were inserted into the multiple cloning sites of pVITRO2 to generate the eukaryotic co-expression plasmid pVITRO2-Mx-NA. The recombinant plasmid was confirmed by restriction endonuclease treatment and sequencing, and it was transfected into the mouse fibroblasts (NIH-3T3) cells. The expression of genes in pVITRO2-Mx-NA were measured by RT-PCR and indirect immunofluorescence assay (IFA). The recombinant plasmid was transfected into CEF cells then RT-PCR and the micro-cell inhibition tests were used to test the antiviral activity for NDV. Our results showed that co-expression vector pVITRO2-Mx-NA was constructed successfully; the expression of Mx and NA could be detected in both NIH-3T3 and CEF cells. The recombinant proteins of Mx and NA protect CEF cells from NDV infection until after 72 h of incubation but the individually mutagenic Mx protein or NA protein protects CEF cells from NDV infection till 48 h post-infection, and co-transfection group decreased significantly NDV infection compared with single-gene transfection group (P<0. 05), indicating that Mx-NA jointing contributed to delaying the infection of NDV in single-cell level and the co-transfection of the jointed genes was more powerful than single one due to their synergistic effects.
authors: .Institute of Animal Breeding and Genetics, Provincial Key Laboratory of Molecular Design/College of Animal Science and Technology, Yangzhou University, Yangzhou, China.
authors: Yani
authors: Y
authors: Zhang
authors: Dezhi
authors: D
authors: Fu
authors: Hao
authors: H
authors: Chen
authors: Zhentao
authors: Z
authors: Zhang
authors: Qingqing
authors: Q
authors: Shi
authors: Ahmed Kamel
authors: AK
authors: Elsayed
authors: Bichun
authors: B
authors: Li
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000998:Antiviral Agents / Q000494:pharmacology*; D048430:Cell Shape / Q000187:drug effects; D002645:Chickens / Q000821:virology*; D005347:Fibroblasts / Q000187:drug effects / Q000473:pathology / Q000821:virology; D005455:Fluorescent Antibody Technique; D015870:Gene Expression / Q000187:drug effects; D051379:Mice; D064547:Myxovirus Resistance Proteins / Q000235:genetics / Q000378:metabolism*; D041681:NIH 3T3 Cells; D009439:Neuraminidase / Q000235:genetics* / Q000378:metabolism; D009522:Newcastle disease virus / Q000187:drug effects* / Q000276:immunology; D010957:Plasmids / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000494:pharmacology*; D014162:Transfection
chemical_list: D000914:Antibodies, Viral; D000998:Antiviral Agents; D064547:Myxovirus Resistance Proteins; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D009439:Neuraminidase
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