If you think that ApE doesn't find all of the ClaI sites (or XbaI or BclI) that you KNOW are present in your sequence, turn off the Dam/Dcm methylation on your sequence and try again. In ApE, open the FASTA file, then use the Features menu to open the GFF3 track info.Īnother way to go is to take the gene model (from a gene page), paste it into an ApE window and then select all, make a new feature (Feature menu), and in the edit feature window that appears press the "upper case only" button. You can add the feature tracks by downloading the GFF3 feature track files using the same menu. gff file in the latest ApE.Īlternatively, you can export a genomic region (from the genome viewer) as a FASTA formatted file (using the menu on the upper left). UNCHECK "Include track configuration data". Dump selected features using version 3 Across currently visible region. You can now export genomic regions from Wormbase directly: In the upper right drop-down menu button, select "Download Track Data". Finds translationally silent restriction sitesĬlick here to make a voluntary donation in support of ApE.Aligns two DNA sequences (or any combination of sequence and ABI trace), with the alignment hyperlinked to the original sequence.Finds potential primers matching user criteria (length, Tm, %GC, self/other complementarity).Translates sequences with optional DNA alignment.Graphic maps that show primer binding sites and all interesting sequence features.Sequence to be annotated and visualized in multiple ways quickly and efficiently.Quick searching and highlighting of all available primers that you (or others) have that hybridize to a sequence.Uses custom feature definition libraries, which allow:.Most analysis windows are hyperlinked to their corresponding sequences, including:.Imports DNA Strider format files (simple enzyme, site lists) available from REBASE.Allows users to define new enzymes by name and recognition site.Has user defined enzyme grouping to distiguish eg.Selects sites that cut more often in one sequence than another (for snip-SNP detection or diagnostic digests).Selects sites matching multiple criteria (union/intersection- cut frequency, site type) in all open windows.Sequences in ABI traces can be aligned directly to a reference sequence, with the alignment hyperlinked back to te trace.Draws pre-defined and user-defined DNA ladders.Copy and save graphics as Windows metafiles (MS Windows only).Saves graphics as encapsulated postscript or scalable vector graphics.Connects graphic and text features with hyperlink double click.Creates graphic restriction maps- linear or circular with features indicated.Text map shows DNA sequence, translation, and features as text-based graphics.Directly BLASTs selected sequence at NCBI or wormbase.Highlights and draws graphic maps using feature annotations from genbank and embl files.Saves files as DNA Strider-compatible or Genbank file format.Reads DNA Strider, Fasta, Genbank and EMBL files.Shows translation, Tm, %GC, ORF of selected DNA in real-time.Highlights text using pre-defined and custom feature libraries.Accurately reflects Dam/Dcm blocking of enzyme sites.Highlights restriction sites in the editing window.Runs in Windows (XP, Vista, 7, 8, and 10) and Mac (OS X v10.11 and above).This work presents a model system to rapidly screen effective DNAzymes targeting SARS and provides a basis for potential therapeutic use of DNA enzymes to combat the SARS infection. Our results demonstrated that this DNAzyme could efficiently cleave the SARS-CoV RNA substrate in vitro and inhibit the expression of the SARS-CoV 5′UTR-eGFP fusion RNA in mammalian cells. Advanced options are available to control specific details of the docking process and obtain intermediate results. The smallest set of input data consists of a protein structure and a DNA or RNA structure in PDB format. A vector containing a fused 5′UTR and enhanced green fluorescent protein (eGFP) was co-transfected with the DNAzyme into E6 cells and the cells expressing eGFP were visualized with fluorescence microscopy and analyzed by fluorescence-activated cell sorting (FACS). The NPDock server provides a user-friendly interface and 3D visualization of the results. In vitro cleavage was performed using an in vitro transcribed 5′UTR RNA substrate. The designed DNAzyme, Dz-104, and its mutant version, Dz-104 (mut), as a control consist of 9 + 9 arm sequences with a 10–23 catalytic core. We designed a RNA-cleaving 10–23 DNAzyme targeting at the loop region of the 5′UTR of SARS-CoV. SARS-CoV total RNA was isolated, extracted from the SARS-CoV-WHU strain and converted into cDNA. A mono-DNA enzyme (Dz-104) possessing the 10–23 catalytic motif was synthesized and tested both in vitro and in cell culture. We report the use of DNAzyme (catalytic DNA) to target the 5′-untranslated region (5′UTR) of a highly conserved fragment in the SARS genome as an approach to suppression of SARS-CoV replication. The worldwide epidemic of severe acute respiratory syndrome (SARS) in 2003 was caused by a novel coronavirus called SARS-CoV.
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