End Cap Structure Research Articles

<em>In vitro</em> Transcription and Capping of <em>Gaussia</em> Luciferase mRNA Followed by HeLa Cell Transfection

In vitro transcription is the synthesis of RNA transcripts by RNA polymerase from a linear DNA template containing the corresponding promoter sequence (T7, T3, SP6) and the gene to be transcribed (Figure 1A). A typical transcription reaction consists of the template DNA, RNA polymerase, ribonucleotide triphosphates, RNase inhibitor and buffer containing Mg(2+) ions. Large amounts of high quality RNA are often required for a variety of applications. Use of in vitro transcription has been reported for RNA structure and function studies such as splicing(1), RNAi experiments in mammalian cells(2), antisense RNA amplification by the "Eberwine method"(3), microarray analysis(4) and for RNA vaccine studies(5). The technique can also be used for producing radiolabeled and dye labeled probes(6). Warren, et al. recently reported reprogramming of human cells by transfection with in vitro transcribed capped RNA(7). The T7 High Yield RNA Synthesis Kit from New England Biolabs has been designed to synthesize up to 180 μg RNA per 20 μl reaction. RNA of length up to 10kb has been successfully transcribed using this kit. Linearized plasmid DNA, PCR products and synthetic DNA oligonucleotides can be used as templates for transcription as long as they have the T7 promoter sequence upstream of the gene to be transcribed. Addition of a 5' end cap structure to the RNA is an important process in eukaryotes. It is essential for RNA stability(8), efficient translation(9), nuclear transport(10) and splicing(11). The process involves addition of a 7-methylguanosine cap at the 5' triphosphate end of the RNA. RNA capping can be carried out post-transcriptionally using capping enzymes or co-transcriptionally using cap analogs. In the enzymatic method, the mRNA is capped using the Vaccinia virus capping enzyme(12,13). The enzyme adds on a 7-methylguanosine cap at the 5' end of the RNA using GTP and S-adenosyl methionine as donors (cap 0 structure). Both methods yield functionally active capped RNA suitable for transfection or other applications(14) such as generating viral genomic RNA for reverse-genetic systems(15) and crystallographic studies of cap binding proteins such as eIF4E(16). In the method described below, the T7 High Yield RNA Synthesis Kit from NEB is used to synthesize capped and uncapped RNA transcripts of Gaussia luciferase (GLuc) and Cypridina luciferase (CLuc). A portion of the uncapped GLuc RNA is capped using the Vaccinia Capping System (NEB). A linearized plasmid containing the GLuc or CLuc gene and T7 promoter is used as the template DNA. The transcribed RNA is transfected into HeLa cells and cell culture supernatants are assayed for luciferase activity. Capped CLuc RNA is used as the internal control to normalize GLuc expression.

Field emission pattern of carbon nanotubes and phase distribution of wave function in the end cap structure

The characteristic pattern of rings seen in field electron emission from capped carbon nanotubes is discussed. The states expected for the p-electrons on a pentagonal ring of carbon atoms are related to their angular periodicity, hence emission from states with a phase change around the ring is less attenuated than that from states with zero phase change. Solution of Schrödinger’s equation for an emitting ring in a system with planar cathode and anode electrodes shows that the wave function at the anode is zero on the axis for any nonzero variation of the phase of emission with azimuthal angle. These considerations suggest that the observed dark centers of rings are due to emission principally from states in the nanotube cap that have nonzero periodicity around the pentagons.

The laser alignment system for the CMS silicon microstrip tracker

In the CMS Microstrip Tracker the algorithms that are used for the online track reconstruction, require the alignment of the tracker modules on a level of better than 100 μ m . A Laser Alignment System has been developed that is able to detect possible movements or deformations of the Tracker mechanical structure with this level of precision. This system works with infrared laser beams which are detected by the silicon microstrip sensors, allowing to monitor movements of a subset of the tracker modules with respect to each other. In this article an overview of the Laser Alignment System is given, and first measurements with the integrated system are presented. The system has been shown to work properly and it confirms the expected mounting precision of the sensor modules in the detector endcaps. During cooling cycles no significant deformation of the endcap structure was observed.

The CMS silicon strip tracker

With more than 15,000 silicon strip modules and an active silicon area of 200 m 2, the Compact Muon Solenoid silicon strip tracker will be the largest silicon tracker ever built. While module mass production has started in 2004, the detector construction has recently entered its crucial phase with modules being assembled onto larger substructures, which in turn are being integrated into the tracker barrel and end-cap structures. In this presentation the detector design will be introduced. The challenges and experiences of the silicon module mass production, with focus on the key components such as sensors and hybrids, will be presented. The status of the integration of modules onto the detector substructures, as well as the construction and integration of the large barrel and end-cap structures will be described. Finally, an overview will be given on the excellent performance of subsystems of the tracker as demonstrated by system tests and test beam experiments.

The CMS silicon strip end cap tracker: Overview and status

While module mass production has started in 2004, the detector construction has entered its hot phase this Spring. The silicon modules are assembled onto substructures, which in turn are integrated into the end cap structures. The detector design, the challenges and experiences of a silicon module mass production of unprecedented scale, the status of the integration of modules onto the detector substructures are introduced as well as the construction and integration of the end cap structures. The excellent performance of the substructures as observed in system tests and test beam experiments are presented.

Dynamic Characteristics of Smart Hull Structures Featuring Piezoelectric Materials

In this paper, dynamic characteristics of an end-capped hull structure with surface bonded piezoelectric actuators are studied. Finite element technique is used to ensure application to practical geometry and boundary conditions of smart hull structure. Modal analysis is conducted to investigate the dynamic characteristics of the hull structure. Piezoelectric self-sensing actuators are attached where the maximum control performance can be obtained. Active controller based on Linear Quadratic Gaussian (LQG) theory is designed to suppress vibration of smart hull structure. It is observed that closed loop damping can be improved with suitable weighting factors in the developed LQG controller.

End cap nucleation of carbon nanotubes

An easily applied graphical approach for facilitating precise tailoring during computational construction of model uncapped or capped single-wall nanotubes or fullerenes is delineated and utilized in this paper. The main enabling concepts are the commonly suggested growth mechanisms for single- and multi-walled carbon nanotubes mediated by end cap structures. The construction protocol described herein can be used to rapidly create any type of armchair, zigzag, chiral or nonchiral defect-free nanotube. Any desired, feasible combination of length and diameter, along with specific placement of hexagonal and pentagonal rings or end caps, can also be controlled. A classification system for end caps is also developed in order to help simplify choosing or describing the particular carbon system under investigation. The suggested methodology is used to systematically model heats of formation of a variety of carbon nanotubes and related fullerenes using AM1 semiempirical calculations. The main factors affecting the calculated physical properties, other than size, are the structures of the various base and terminating end caps. Finally, we comment on the possible relationship of the construction methodology to mechanisms for carbon nanotube nucleation.

Fullerene negative ion irradiation toward double-walled carbon nanotubes using low energy magnetized plasma

We here present the details of fullerene filling inside double-walled (DWNTs) and multi-walled carbon nanotubes (MWNTs) and also demonstrate plasma ion-irradiation effects on various kinds of nanotube structures. It is found that both the degree of the structural deformation and the fullerene filling efficiency are directly proportional to the applied bias voltage and plasma density used. In particular, in the case of relatively low substrate-bias application, irradiated fullerenes are only adsorbed on the surroundings of the DW/MWNTs, which are kept under end-cap structure. However, it should be noted that fullerene-like material filled DWNTs are shown to be formed by relatively high substrate-bias application, which frequently provokes nanotubes to form open-end structures.

Position dependence of the rous sarcoma virus negative regulator of splicing element reflects proximity to a 5′ splice site

Rous sarcoma virus (RSV) requires incomplete splicing of its viral transcripts to maintain efficient replication. A splicing inhibitor element, the negative regulator of splicing (NRS), is located near the 5′ end of the RNA but the significance of this positioning is not known. In a heterologous intron the NRS functions optimally when positioned close to the authentic 5′ splice site. This observation led us to investigate the basis of the position dependence. Four explanations were put forth and stressed the role of three major elements involved in splicing, the 3′ splice site, the 5′ splice site, and the 5′ end cap structure. NRS function was unrelated to its position relative to the 3′ splice site or the cap structure and appeared to depend on its position relative to the authentic 5′ splice site. We conclude that position dependence may reflect distance constraints necessary for competition of the NRS with the authentic 5′ splice site for pairing with the 3′ splice sites.

Relationship between the end-cap structure of polycarbonates and their impact resistance

Using various phenols as the end-capping agent, polycarbonates (PCs) with the same degrees of polymerization were prepared by phosgenation of bisphenol A. The end-capping agents greatly affected the fluidity, thermal resistance and impact resistance of the PCs. It was found that, compared to commercially available PCs end-capped with 4-tert-butylphenol, the PCs end-capped with 4-tert-octylphenol or 4-α-cumylphenol were superior in impact strength with a better balance of fluidity, thermal resistance and impact resistance. From the results of dynamic mechanical measurements, improvement in performance is explained by the high mobility of my prepared PCs as compared to commercial ones, that is, energy dissipation occurs significantly in the polymer molecule, and contributes to the improvement in impact resistance.

Translational control and synaptic activity: a role for IRES elements?

Initiation of translation of cellular mRNAs proceeds mainly through the recruitment of the ribosome via the 5’-end cap structure, followed by a linear scanning of the mRNA leader sequence and positioning on the initiator codon. An alternative (cap-independent) mechanism involves the direct binding of the translational apparatus within the mRNA via cis-elements called internal ribosome entry sites (IRESs). Initially detected in some viral mRNAs, IRES elements are found in a restricted number of cellular mRNAs and are thought to facilitate the recruitment of the ribosome in conditions with reduced availability of the eukaryotic initiation factor (eIF) 4F cap-binding complex, such as apoptosis or mitosis.

IRES elements: fine-tuning of gene expression at the translational level

Some viral mRNAs contain internal ribosome entry sites (IRESs), which are cis elements that allow the binding of the translational apparatus directly within the mRNA. IRESs therefore offer an alternative to the general mechanism of the ribosome scanning from the 5′-end cap structure to the initiator codon that prevails for most cellular mRNAs. Interestingly, IRESs have also been identified in a few cellular messengers. Whereas viral IRESs provide the advantage of initiating translation in the case of uncapped mRNAs (e.g. picornaviruses mRNAs) or under conditions where the global host protein synthesis is inhibited (e.g. during infection), the biological function of cellular IRESs is still unclear. Recent ex vivo data have shown that certain cellular IRESs could mediate a cell-cycle-dependent translational control. In the July issue of the Journal of Cell Biology, Creancier and co-workers describe the first in vivo study of the activity of a mammalian cellular IRES, the human fibroblast growth factor 2 (FGF2) IRES (Ref. 1xFibroblast growth factor 2 internal ribosome entry site (IRES) activity ex vivo and in transgenic mice reveals a stringent tissue-specific regulation. Creancier, L. et al. J. Cell Biol. 2000; 150: 275–281Crossref | PubMed | Scopus (103)See all ReferencesRef. 1).The authors have created transgenic mice that express a dicistronic mRNA bearing two open reading frames (ORFs) encoding two distinct luciferases. In this standard assay, the upstream ORF is translated via the cap-dependent scanning mechanism, whereas translation of the second ORF is driven by the IRES located between them. By this method they could measure the IRES activity of the FGF2 mRNA specifically. Strikingly, they observed a stage- and tissue-specific modulation of the FGF2 IRES activity. It was high in the embryo (especially in the brain and the heart at E16) in comparison with the adult, in which this strong IRES activity appeared to be restricted mainly to the brain. This contrasted with the widespread strong activity of picornavirus IRESs. How does the tropism of FGF2 IRES activity fit with the broad expression of this growth factor all over the organism? The fact that FGF2 translation can be initiated via the IRES but also by a cap-dependent mechanism could explain such a discrepancy. Nevertheless, the poor IRES activity in organs that otherwise express FGF2 raises the question of the inducibility of IRESs. Indeed, stimulation of the FGF2 IRES could represent a means to adapt the level of FGF2 expression selectively in response to a given stimulus. This could involve potential as yet unidentified trans-acting factors that would be active in the adult brain while silent in other organs (in the steady state).The stringent in vivo regulation of the FGF2 IRES activity described in this paper emphasizes the role of translation in the control of gene expression, and the strategy applied here can be used for the in vivo study of other cellular IRESs. This could allow to address the question whether IRES elements coordinately regulate the translation of a subclass of mRNAs. Furthermore, the understanding of the molecular mechanisms underlying IRES regulation in vivo might lead to strategies designed to control the expression of endogenous factors artificially, as well as to target the expression of trans genes of interest. The animal model described here will represent an indispensable tool to validate such strategies.

A 54-kDa Fragment of the Poly(A)-specific Ribonuclease Is an Oligomeric, Processive, and Cap-interacting Poly(A)-specific 3′ Exonuclease

We have previously identified a HeLa cell 3' exonuclease specific for degrading poly(A) tails of mRNAs. Here we report on the purification and identification of a calf thymus 54-kDa polypeptide associated with a similar 3' exonuclease activity. The 54-kDa polypeptide was shown to be a fragment of the poly(A)-specific ribonuclease 74-kDa polypeptide. The native molecular mass of the nuclease activity was estimated to be 180-220 kDa. Protein/protein cross-linking revealed an oligomeric structure, most likely consisting of three subunits. The purified nuclease activity released 5'-AMP as the reaction product and degraded poly(A) in a highly processive fashion. The activity required monovalent cations and was dependent on divalent metal ions. The RNA substrate requirement was investigated, and it was found that the nuclease was highly poly(A)-specific and that only 3' end-located poly(A) was degraded by the activity. RNA substrates capped with m(7)G(5')ppp(5')G were more efficiently degraded than noncapped RNA substrates. Addition of free m(7)G(5')ppp(5')G cap analogue inhibited poly(A) degradation in vitro, suggesting a functional link between the RNA 5' end cap structure and poly(A) degradation at the 3' end of the RNA.

Evidence that the 5'-end cap structure is essential for encapsidation of hepatitis B virus pregenomic RNA.

Hepatitis B virus (HBV) replicates by reverse transcription of an RNA intermediate, the pregenomic RNA. The first step of HBV genome replication is the encapsidation of the pregenomic RNA encoding the encapsidation signal, termed epsilon, into the core particles, which is preceded by recognition and binding of HBV DNA polymerase to epsilon. The pregenomic RNA contains two identical epsilon elements due to its terminal redundancy: one near the 5' end and another near the 3' end. Despite the fact that both epsilon elements have an identical sequence, only the 5' epsilon, but not the 3' epsilon, is functional for encapsidation. To understand the molecular nature of this position effect, we made a series of lacZ RNA expression plasmids which contain the epsilon element at various positions from the 5' end of the transcripts. Following transfection, the lacZ RNAs in cytoplasmic core particles were measured by RNase protection assay for encapsidation. The results indicated that the lacZ RNAs with epsilon positioned up to 65 nucleotides from the 5' end were encapsidated, whereas the lacZ RNAs with epsilon positioned further downstream were not. Interestingly, the cap-free lacZ RNA transcribed by T7 RNA polymerase was not encapsidated, implying that the 5' cap structure is required for encapsidation of the pregenomic RNA. We hypothesized that HBV DNA polymerase must somehow recognize the cap structure and/or its associated factors, as well as the 5' epsilon, for encapsidation to occur.

Human histamine H2 receptor gene: multiple transcription initiation and tissue-specific expression

We have characterized the genomic structure of 12.3 kb of the 5′-flanking region of the human histamine H2 receptor gene. The multiple transcription initiation sites of the human histamine H2 receptor gene were mapped utilizing the 5′-end cap structure of mRNA. We found that a 85 bp segment (−610–−525 bp) immediately upstream of the initiation site exhibits a strong promoter activity in the gastric adenocarcinoma, MNK45, expressing human histamine H2 receptor. A 4.8 kb transcript of the human histamine H2 receptor gene was found in the placenta, spinal cord, lymph node and bone marrow in addition to the previously reported tissues including the heart, brain and stomach, whereas a 1.8 kb transcript was observed in almost all tissues examined. 3′-rapid amplification of cDNA ends revealed the corresponding length of the 3′-untranslated region. These results suggest that the 3′-untranslated region may be involved in the differential expression.

Recombinant-baculovirus-expressed PB 2 subunit of the influenza A virus RNA polymerase binds cap groups as an isolated subunit

The influenza A virus RNA-dependent RNA polymerase catalyzes several reactions in transcription and replication of the genome RNA. The first step in viral mRNA synthesis is the recognition of the 5′ end cap structure of host cell hnRNA and the cleavage of the RNA substrate between 10 and 14 nucleotides from the 5′ end to generate capped primers for initiation of transcription of virus-specific mRNAs. This report describes the use of an in vitro UV crosslinking and protein renaturation assay to identify the polymerase subunits which interact with the 5′ end cap structure of an artificial RNA substrate. Our results showed, for the first time, that purified polymerase subunit PB 2 expressed by recombinant baculovirus in insect cells possessed cap-binding activity by itself after renaturation by Escherichia coli thioredoxin, whereas cleavage of the artificial capped substrate required the holoenzyme expressed in insect cells triply-infected with baculovirus containing all three polypeptide components, PB 1, PB 2, and PA. Purified polyclonal anti-PB 2 IgG inhibited the binding activity; anti-PB 1 and anti-PA IgGs did not.

Synthesis and surfactant properties of novel acrylic acid co-oligomers containing fluoroalkylated end-groups: a new approach to polymeric inhibitors of human immunodeficiency virus type-1

Reactions of fluoroalkanoyl peroxides with acrylic acid, and with alkyl methacrylates or trimethylvinylsilane, give a series of new fluoroalkylated acrylic acid/alkyl methacrylate and /trimethylvinylsilane co-oligomers in excellent to moderate yield under very mild conditions. Fluoroalkylated acrylic acid/alkyl methacrylate co-oligomers are soluble in common organic solvents such as methanol, DMSO and tetrahydrofuran, and slightly soluble in non-polar aromatic solvents such as benzene, toluene and xylene. However, these co-oligomers do not have good solubility in both water and aromatic solvents. In contrast, fluoroakylated acrylic acid/trimethylvinylsilane co-oligomers are readily soluble in not only in water but also in common organic solvent such as methanol, ethanol, tetrahydrofuran, carbon tetrachloride, chloroform, DMF, DMSO, benzene, toluene and xylene. This feature is due to the trimethylsilyl side-chains possessing good oleophilic properties. Fluoroalkylated co-oligomers, in particular fluoroalkylated acrylic acid/trimethylvinylsilane co-oligomers, reduce the surface tension of both water and m-xylene effectively to around 15–20 mN m −1, the same as for low-molecular weight fluorinated surfactants, though the co-oligomers have high molecular weights. These fluoroalkylated silicon-containing co-oligomers exhibit a clear break point resembling a CMC, which suggest that they form intra- or inter-molecular aggregations in aqueous solutions owing to their unique structure (fluoroalkylated end-capped structure). The fluoroalkylated acrylic acid co-oligomers inhibit HIV-1 induced cytopathogenesis. Relative to dextran sulfate, superior anti-HIV-1 activity could be obtained with fluoroalkylated acrylic acid/trimethylvinylsilane co-oligomers; however, fluoroalkylated acrylic acid cooligomers containing hydroxy groups were inactive. An obvious correlation exists between anti-HIV-1 activity and the hydrophilic or lipophilic property of these co-oligomers, i.e. they provide a most adequate HLB value for anti-HIV-1 activity. More hydrophilic or more oleophilic fluoroalkylated co-oligomers were less active (or inactive), suggesting that more hydrophilic or more oleophilic fluoroalkylated co-oligomers have a weaker effect on the interaction between gp/120 (or gp41) in HIV-1 and CD4 receptors. In fluoroalkylated silicon co-oligomers, as the degree of the reduction in surface tension of water becomes higher the anti-HIV-1 activity becomes higher, suggesting that the more adsorbable co-oligomers at the water/air interface have a higher anti-HIV-1 activity. A new correlation between anti-HIV-1 activity and properties of fluoroalkylated acrylic acid co-oligomers is indicated, affording a useful new strategy for exploring potent new polymeric inhibitors of HIV-1.

Creation of deletion, insertion and substitution mutations using a single pair of primers and PCR.

BioTechniquesVol. 20, No. 2 BenchmarksOpen AccessCreation of Deletion, Insertion and Substitution Mutations Using a Single Pair of Primers and PCRMartin John Glenton Hughes & David William AndrewsMartin John Glenton HughesMcMaster University, Hamilton, ON, CanadaSearch for more papers by this author & David William Andrews*Address correspondence to David W. Andrews, McMaster University, Department of Biochemistry, 1200 Main Street West, Hamilton, ON L8N 3Z5, Canada.McMaster University, Hamilton, ON, CanadaSearch for more papers by this authorPublished Online:2 Aug 2018https://doi.org/10.2144/96202bm06AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinkedInRedditEmail FiguresReferencesRelatedDetailsCited ByA Syringe‐Based DNAzyme Sensor for Bacterial Detection7 April 2021 | Analysis & Sensing, Vol. 1, No. 2In Vitro Mutagenesis Using Double-Stranded DNA Templates: Selection of Mutants with DpnI1 March 2018 | Cold Spring Harbor Protocols, Vol. 2018, No. 3A Mutant dec-1 Transgene Induces Dominant Female Sterility in Drosophila melanogaster1 November 2007 | Genetics, Vol. 177, No. 3Genomic structure and functional characterization of the promoter region of human IκB kinase-related kinase IKKi/IKKɛ geneGene, Vol. 353, No. 1The N-terminal prodomain of sV23 is essential for the assembly of a functional vitelline membrane network in DrosophilaDevelopmental Biology, Vol. 270, No. 1A Direct Binding Site for Grb2 Contributes to Transformation and Leukemogenesis by the Tel-Abl (ETV6-Abl) Tyrosine KinaseMolecular and Cellular Biology, Vol. 24, No. 11Active site residues and mechanism of UDP-glucose dehydrogenase14 December 2003 | European Journal of Biochemistry, Vol. 271, No. 1Mutational Analysis of the TonB1 Energy Coupler of Pseudomonas aeruginosaJournal of Bacteriology, Vol. 184, No. 6The Functions of the Multiproduct and Rapidly Evolving dec-1 Eggshell Gene Are Conserved Between Evolutionarily Distant Species of Drosophila1 November 2001 | Genetics, Vol. 159, No. 3gmhX, a novel gene required for the incorporation of l-glycero-d -manno-heptose into lipooligosaccharide in Neisseria meningitidis The GenBank accession number for the sequence of the ice-2 operon in Neisseria meningitidis NMB is AF036242.Microbiology, Vol. 147, No. 8Nucleotide-dependent Binding of the GTPase Domain of the Signal Recognition Particle Receptor β-Subunit to the α-SubunitJournal of Biological Chemistry, Vol. 275, No. 35Spatial and Temporal Expression of a Polysphondylium Spore-Specific GeneDevelopmental Biology, Vol. 224, No. 1Evidence that the 5′-End Cap Structure Is Essential for Encapsidation of Hepatitis B Virus Pregenomic RNAJournal of Virology, Vol. 74, No. 12Modulation of casein proteolysis by lactococcal peptidase gene inactivationInternational Dairy Journal, Vol. 10, No. 9A Site-specific, Membrane-dependent Cleavage Event Defines the Membrane Binding Domain of FtsYJournal of Biological Chemistry, Vol. 274, No. 47Dominant Negative Mutants Implicate STAT5 in Myeloid Cell Proliferation and Neutrophil DifferentiationBlood, Vol. 93, No. 12Multiple lysophosphatidic acid acyltransferases in Neisseria meningitidisMolecular Microbiology, Vol. 32, No. 5Improved Protocol for High-Throughput Cysteine Scanning MutagenesisStefan Howorka & Hagan Bayley15 August 2018 | BioTechniques, Vol. 25, No. 5Endothelin Converting-Enzyme-1 mRNA Expression in Human Cardiovascular Disease3 July 2009 | Clinical and Experimental Hypertension, Vol. 20, No. 4Modification of a PCRBased Site-Directed Mutagenesis MethodConstance L. Fisher & Guo Kui Pei7 August 2018 | BioTechniques, Vol. 23, No. 4Transcriptional Induction of Collagenase-1 in Differentiated Monocyte-like (U937) Cells is Regulated by AP-1 and an Upstream C/EBP-β SiteJournal of Biological Chemistry, Vol. 272, No. 18Primer-Directed Mutagenesis of an Intact Plasmid by Using Pwo DNA Polymerase in Long Distance Inverse PCRRachmat Hidajat & Patricia McNicol7 August 2018 | BioTechniques, Vol. 22, No. 1Site-Directed Mutagenesis for Large Insertions by Oligonucleotide Primers in Optimized Molar RatiosDong Wang & Hei Sook Sul7 August 2018 | BioTechniques, Vol. 22, No. 1 Vol. 20, No. 2 Follow us on social media for the latest updates Metrics Downloaded 381 times History Published online 2 August 2018 Published in print February 1996 Information© 2018 Author(s)PDF download

Identification of the 3′ - and 5′ -ends of beet necrotic yellow vein virus RNAs

Beet necrotic yellow vein virus RNAs are characterized by a 5′ -end cap structure and polyadenylated 3′ -ends. The presence of a poly(A) sequence with major lengths ranging from 65–140 nucleotides could be demonstrated by homochromatography and sequencing methods. Beet necrotic yellow vein virus RNAs could be 5′ -end labelled with kinase only after enzymatic or chemical cap elimination followed by dephosphorylation. The 5′ -end structure is found to be of cap O type: m 7GpppNp.