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10.12688/wellcomeopenres.16322.1 http://scihub22266oqcxt.onion/10.12688/wellcomeopenres.16322.1 33313418!7721065!33313418     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=33313418&cmd=llinks): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 215 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Wellcome+Open+Res 2020 ; 5 (ä): 261 Nephropedia Template TP {{tp|p=33313418|t=2020. Integrative vectors for regulated expression of SARS-CoV-2 proteins implicated in RNA metabolism |pdf=|usr=}}{{33313418}} gab.com Text Integrative vectors for regulated expression of SARS-CoV-2 proteins implicated in RNA metabolism JO: Wellcome Open Res http://www.kidney.de/pget.php?&tw=1&pmid=33313418 #FOAvip Infection with SARS-CoV-2 is expected to result in substantial reorganization of host cell RNA metabolism. We identified 14 proteins that were predicted to interact with host RNAs or RNA binding proteins, based on published data for SARS-CoV and SARS-CoV-2. Here, we describe a series of affinity-tagged and codon-optimized expression constructs for each of these 14 proteins. Each viral gene was separately tagged at the N-terminus with Flag-His (8), the C-terminus with His (8)-Flag, or left untagged. The resulting constructs were stably integrated into the HEK293 Flp-In T-REx genome. Each viral gene was expressed under the control of an inducible Tet-On promoter, allowing expression levels to be tuned to match physiological conditions during infection. Expression time courses were successfully generated for most of the fusion proteins and quantified by western blot. A few fusion proteins were poorly expressed, whereas others, including Nsp1, Nsp12, and N protein, were toxic unless care was taken to minimize background expression. All plasmids can be obtained from Addgene and cell lines are available. We anticipate that availability of these resources will facilitate a more detailed understanding of coronavirus molecular biology. Twit Text FOAVip Integrative vectors for regulated expression of SARS-CoV-2 proteins implicated in RNA metabolism ????Wellcome Open Res http://www.kidney.de/pget.php?&tw=1&pmid=33313418 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33313418 #FOAvip English Wikipedia <ref>{{Cite pmid|33313418}}</ref> Integrative vectors for regulated expression of SARS-CoV-2 proteins implicated in RNA metabolism #MMPMID33313418 Bresson S; Robertson N; Sani E; Turowski TW; Shchepachev V; Kompauerova M; Spanos C; Helwak A; Tollervey D Wellcome Open Res 2020[]; 5 (ä): 261 PMID33313418show ga Infection with SARS-CoV-2 is expected to result in substantial reorganization of host cell RNA metabolism. We identified 14 proteins that were predicted to interact with host RNAs or RNA binding proteins, based on published data for SARS-CoV and SARS-CoV-2. Here, we describe a series of affinity-tagged and codon-optimized expression constructs for each of these 14 proteins. Each viral gene was separately tagged at the N-terminus with Flag-His (8), the C-terminus with His (8)-Flag, or left untagged. The resulting constructs were stably integrated into the HEK293 Flp-In T-REx genome. Each viral gene was expressed under the control of an inducible Tet-On promoter, allowing expression levels to be tuned to match physiological conditions during infection. Expression time courses were successfully generated for most of the fusion proteins and quantified by western blot. A few fusion proteins were poorly expressed, whereas others, including Nsp1, Nsp12, and N protein, were toxic unless care was taken to minimize background expression. All plasmids can be obtained from Addgene and cell lines are available. We anticipate that availability of these resources will facilitate a more detailed understanding of coronavirus molecular biology. äIntegrative vectors for regulated expression of SARS-CoV-2 proteins im(epmc).pdf DeepDyve Pubget Overpricing