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10.1002/jgm.640 http://scihub22266oqcxt.onion/10.1002/jgm.640 15543523!7166999!15543523     free     free     free Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Gene+Med 2005 ; 7 (1): 97-107 Nephropedia Template TP {{tp|p=15543523|t=2005. Antisense downregulation of SARS-CoV gene expression in Vero E6 cells |pdf=|usr=}}{{15543523}} gab.com Text Antisense downregulation of SARS-CoV gene expression in Vero E6 cells JO: J Gene Med http://www.kidney.de/pget.php?&tw=1&pmid=15543523 #FOAvip BACKGROUND: Severe acute respiratory syndrome (SARS) is caused by a novel coronavirus (SARS-CoV). It is an enveloped, single-stranded, plus-sense RNA virus with a genome of approximately 30 kb. The structural proteins E, M and N of SARS-CoV play important roles during host cell entry and viral morphogenesis and release. Therefore, we have studied whether expression of these structural proteins can be down-regulated using an antisense technique. METHODS: Vero E6 cells were transfected with plasmid constructs containing exons of the SARS-CoV structural protein E, M or N genes or their exons in frame with the reporter protein EGFP. The transfected cell cultures were treated with antisense phosphorothioated oligonucleotides (antisense PS-ODN, 20mer) or a control oligonucleotide by addition to the culture medium. RESULTS: Among a total of 26 antisense PS-ODNs targeting E, M and N genes, we obtained six antisense PS-ODNs which could sequence-specifically reduce target genes expression by over 90% at the concentration of 50 microM in the cell culture medium tested by RT-PCR. The antisense effect was further proved by down-regulating the expression of the fusion proteins containing the structural proteins E, M or N in frame with the reporter protein EGFP. In Vero E6 cells, the antisense effect was dependent on the concentrations of the antisense PS-ODNs in a range of 0-10 microM or 0-30 microM. CONCLUSIONS: The antisense PS-ODNs are effective in downregulation of SARS. The findings indicate that antisense knockdown of SARS could be a useful strategy for treatment of SARS, and could also be suitable for studies of the pathological function of SARS genes in a cellular model system. Twit Text FOAVip Antisense downregulation of SARS-CoV gene expression in Vero E6 cells ????J Gene Med http://www.kidney.de/pget.php?&tw=1&pmid=15543523 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=15543523 #FOAvip English Wikipedia <ref>{{Cite pmid|15543523}}</ref> Antisense downregulation of SARS-CoV gene expression in Vero E6 cells #MMPMID15543523 Shi Y; Luo H; Jia J; Xiong J; Yang D; Huang B; Jin Y J Gene Med 2005[Jan]; 7 (1): 97-107 PMID15543523show ga BACKGROUND: Severe acute respiratory syndrome (SARS) is caused by a novel coronavirus (SARS-CoV). It is an enveloped, single-stranded, plus-sense RNA virus with a genome of approximately 30 kb. The structural proteins E, M and N of SARS-CoV play important roles during host cell entry and viral morphogenesis and release. Therefore, we have studied whether expression of these structural proteins can be down-regulated using an antisense technique. METHODS: Vero E6 cells were transfected with plasmid constructs containing exons of the SARS-CoV structural protein E, M or N genes or their exons in frame with the reporter protein EGFP. The transfected cell cultures were treated with antisense phosphorothioated oligonucleotides (antisense PS-ODN, 20mer) or a control oligonucleotide by addition to the culture medium. RESULTS: Among a total of 26 antisense PS-ODNs targeting E, M and N genes, we obtained six antisense PS-ODNs which could sequence-specifically reduce target genes expression by over 90% at the concentration of 50 microM in the cell culture medium tested by RT-PCR. The antisense effect was further proved by down-regulating the expression of the fusion proteins containing the structural proteins E, M or N in frame with the reporter protein EGFP. In Vero E6 cells, the antisense effect was dependent on the concentrations of the antisense PS-ODNs in a range of 0-10 microM or 0-30 microM. CONCLUSIONS: The antisense PS-ODNs are effective in downregulation of SARS. The findings indicate that antisense knockdown of SARS could be a useful strategy for treatment of SARS, and could also be suitable for studies of the pathological function of SARS genes in a cellular model system. |*Down-Regulation[MESH] |*Gene Expression Regulation, Viral[MESH] |Animals[MESH] |Chlorocebus aethiops[MESH] |Dose-Response Relationship, Drug[MESH] |Green Fluorescent Proteins/metabolism[MESH] |Luminescent Proteins/*genetics[MESH] |Microscopy, Confocal[MESH] |Oligonucleotides, Antisense/chemistry/genetics/metabolism/*pharmacology[MESH] |Plasmids[MESH] |Recombinant Fusion Proteins/metabolism[MESH] |Reverse Transcriptase Polymerase Chain Reaction[MESH] |Severe acute respiratory syndrome-related coronavirus/genetics/*metabolism[MESH]Antisense downregulation of SARS-CoV gene expression in Vero E6 cells (epmc).pdf DeepDyve Pubget Overpricing