Effects of sequences of prokaryotic origin on titer and transgene expression in retroviral vectors.


:Transcriptionally targeted MLV-based ProCon vectors allow expression of the transduced gene in a promoter-specific manner by replacement of the viral U3 region with a heterologous promoter. In order to evaluate the effects of sequence elements present in ProCon vectors on transgene expression (enhanced green fluorescence protein, EGFP), a series of deletion constructs mimicking the situation in proviral DNA following promoter conversion, where expression of the EGFP gene is driven by three different constitutive promoters (MLV U3, mCMV, and hCMV) in the context of a 5'LTR, respectively, were generated and tested in transient transfection experiments. We discovered that modifications in the 3'LTR have only marginal effects on the EGFP expression and the sequence between the promoter and the transgene did not influence EGFP expression at all. On the other hand, EGFP expression was reduced by up to 17-fold in cells transfected with constructs containing SV40neo and/or pBR322ori sequences. To study this effect in transduced cells, we generated a series of retroviral vectors in which these elements were deleted in various combinations and found that an increase in EGFP expression and viral titer was also consistently obtained using vectors lacking these elements, although this was much smaller than that observed using the expression constructs. A vector containing the gene for puromycin resistance (pac) in place of the neomycin resistance gene (neo) was also tested, and found to result in improved vector titers and transgene expression. We conclude that, where possible, the inclusion of neo and ori sequences in retroviral vectors should be avoided, and that, if selection of infected cells is necessary, the pac, rather than neo gene should be used.






Hlavaty J,Portsmouth D,Stracke A,Salmons B,Günzburg WH,Renner M




Has Abstract


2004-12-05 00:00:00
















  • HIV-1 clones resistant to a small molecule CCR5 inhibitor use the inhibitor-bound form of CCR5 for entry.

    abstract::Human immunodeficiency virus type 1 (HIV-1) infection can be inhibited by small molecules that target the CCR5 coreceptor. Here, we describe some properties of clonal viruses resistant to one such inhibitor, SCH-D, using both chimeric, infectious molecular clones and Env-pseudotypes. Studies using combinations of CCR5...


    pub_type: 杂志文章


    authors: Pugach P,Marozsan AJ,Ketas TJ,Landes EL,Moore JP,Kuhmann SE

    更新日期:2007-04-25 00:00:00

  • Deletion of 55 open reading frames from the termini of vaccinia virus.

    abstract::Each copy of the inverted terminal repeat of vaccinia virus consists of 8 kb of DNA containing 9 ORFS flanked near the terminus of the genome by 4 kb of repetitive DNA which in turn contains blocks of tandem repeats. Using plasmids containing repetitive DNA as the external arm, we have generated deletions at both the ...


    pub_type: 杂志文章


    authors: Perkus ME,Goebel SJ,Davis SW,Johnson GP,Norton EK,Paoletti E

    更新日期:1991-01-01 00:00:00

  • Electron microscopy of binding of Epstein-Barr virus (EBV) nuclear antigen (EBNA-1) to EBV DNA.

    abstract::Specific binding sites for Epstein-Barr virus (EBV) nuclear antigen (EBNA-1), isolated and semipurified from EBV-transformed nonproductive Raji cells, were visualized on the molecule of EBV DNA by electron microscopy and mapped. Two measures had to be applied to counteract the limited purity of the EBNA-1 preparation:...


    pub_type: 杂志文章


    authors: Reischig J,Bartsch D,Polack A,Vonka V,Hirsch I

    更新日期:1987-10-01 00:00:00

  • Epitope exposure on functional, oligomeric HIV-1 gp41 molecules.

    abstract::We have used cells infected with the HIV-1 molecular clone HX10 to study the binding of monoclonal antibodies (mAbs) to different epitopes within the extracellular domain of the HIV-1 transmembrane glycoprotein gp41. Gp41 mAb binding to the infected cells at 4 degrees was variable but weaker than the binding of an ant...


    pub_type: 杂志文章


    authors: Sattentau QJ,Zolla-Pazner S,Poignard P

    更新日期:1995-01-10 00:00:00

  • Identification and characterization of a new base substitution in the vaccine strain of Sabin 3 poliovirus.

    abstract::The complete RNA sequence of Sabin 3 (LED3) used in vaccine in the United States has been determined. The LED3 Sabin 3 sequence contains the attenuating mutations at bases 472 and 2034 but differs from that published by Stanway et al. (Nucleic Acids Res., 11, 5629-5643, 1983) at two other base positions, 2493 and 6061...


    pub_type: 杂志文章


    authors: Weeks-Levy C,Tatem JM,DiMichele SJ,Waterfield W,Georgiu AF,Mento SJ

    更新日期:1991-12-01 00:00:00

  • Evaluation of antibody parameters as potential correlates of protection or enhancement by experimental vaccines to equine infectious anemia virus.

    abstract::We previously demonstrated in trials of a variety of experimental vaccines to equine infectious anemia virus (EIAV) a remarkable spectrum of efficacy ranging from sterilizing protection to severe enhancement of virus replication and disease, depending on the immunization strategy used. This range of vaccine efficacy o...


    pub_type: 杂志文章


    authors: Hammond SA,Raabe ML,Issel CJ,Montelaro RC

    更新日期:1999-09-30 00:00:00

  • The structure of three bacteriophage T4 genes required for tail-tube assembly.

    abstract::Three different protein molecules copurify with T4 tail tubes after the tubes are released from the baseplate by guanidine hydrochloride treatment. These tube-associated proteins (TAPs) are the products of genes 29, 48, and 54. To further investigate the structural roles that these proteins may play in T4 tail assembl...


    pub_type: 杂志文章


    authors: Ishimoto LK,Ishimoto KS,Cascino A,Cipollaro M,Eiserling FA

    更新日期:1988-05-01 00:00:00

  • Functional domains in the feline immunodeficiency virus nucleocapsid protein.

    abstract::Retroviral nucleocapsid (NC) proteins are small Gag-derived products containing one or two zinc finger motifs that mediate genomic RNA packaging into virions. In this study, we addressed the role of the feline immunodeficiency virus (FIV) NC protein in the late stages of virus replication by analyzing the assembly phe...


    pub_type: 杂志文章


    authors: Manrique ML,Rauddi ML,González SA,Affranchino JL

    更新日期:2004-09-15 00:00:00

  • The effect of inhibitors on the growth of the entomopoxvirus from Amsacta moorei in Lymantria dispar (gypsy moth) cells.

    abstract::Within the family of Poxviridae, the entomopoxviruses are the most distant relatives of the more well-known and intensively studied orthopoxviruses (vaccinia and variola). The growth of the entomopoxvirus from Amsacta moorei (AmEPV) has been characterized in cell culture and compared to that of vaccinia virus (VV), th...


    pub_type: 杂志文章


    authors: Winter J,Hall RL,Moyer RW

    更新日期:1995-08-20 00:00:00

  • Regulation of a geminivirus coat protein promoter by AL2 protein (TrAP): evidence for activation and derepression mechanisms.

    abstract::Tomato golden mosaic virus (TGMV) is a bipartite member of the subgroup III Geminiviridae. Like all geminiviruses, TGMV replicates in the nucleus of susceptible cells by rolling circle replication (RCR). Double-stranded replicative form DNA generated during RCR serves as template for the transcription of viral genes b...


    pub_type: 杂志文章


    authors: Sunter G,Bisaro DM

    更新日期:1997-06-09 00:00:00

  • Adenoviral infection induces a multi-faceted innate cellular immune response that is mediated by the toll-like receptor pathway in A549 cells.

    abstract::Adenovirus vectors are known to induce certain genes and impact innate response networks, but a broad understanding of this process and its mechanisms is currently lacking. For this reason, we chose to investigate and characterize Ad innate immunity using homogeneous, primary MEF cells replete with all the elements of...


    pub_type: 杂志文章


    authors: Hartman ZC,Black EP,Amalfitano A

    更新日期:2007-02-20 00:00:00

  • Antibody neutralization of retargeted measles viruses.

    abstract::The measles virus (MV) vaccine lineage is a promising oncolytic but prior exposure to the measles vaccine or wild-type MV strains limits treatment utility due to the presence of anti-measles antibodies. MV entry can be redirected by displaying a polypeptide ligand on the Hemagglutinin (H) C-terminus. We hypothesized t...


    pub_type: 杂志文章


    authors: Lech PJ,Pappoe R,Nakamura T,Tobin GJ,Nara PL,Russell SJ

    更新日期:2014-04-01 00:00:00

  • Chlorella virus NY-2A encodes at least 12 DNA endonuclease/methyltransferase genes.

    abstract::The 380-kb chlorella virus NY-2A genome is highly methylated; 45% of the cytosines are 5-methylcytosine (5mC) and 37% of the adenines are N6-methyladenine (6mA). Based on the sensitivity/resistance of NY-2A DNA to 80 methylation-sensitive DNA restriction endonucleases, the virus is predicted to encode at least 10 DNA ...


    pub_type: 杂志文章


    authors: Zhang Y,Nelson M,Nietfeldt J,Xia Y,Burbank D,Ropp S,Van Etten JL

    更新日期:1998-01-20 00:00:00

  • Location, size, and complexity of epitopes on the coat protein of beet necrotic yellow vein virus studied by means of synthetic overlapping peptides.

    abstract::Five regions on the coat protein of BNYVV which had been shown previously to be involved in the formation of continuous epitopes were further analyzed by means of synthetic overlapping peptides. It was found that at least some of these regions may encompass several overlapping epitopes (or parts thereof). Four monoclo...


    pub_type: 杂志文章


    authors: Commandeur U,Koenig R,Manteuffel R,Torrance L,Lüddecke P,Frank R

    更新日期:1994-01-01 00:00:00

  • Molecular and serological characterization of SARS-CoV-2 infection among COVID-19 patients.

    abstract:BACKGROUND:SARS-CoV-2 is a novel coronavirus and the cause of COVID-19. More than 80% of COVID-19 patients exhibit mild or moderate symptoms. In this study, we investigated the dynamics of viral load and antibodies against SARS-CoV-2 in a longitudinal cohort of COVID-19 patients with severe and mild/moderate diseases. ...


    pub_type: 杂志文章


    authors: Li L,Liang Y,Hu F,Yan H,Li Y,Xie Z,Huang L,Zhao J,Wan Z,Wang H,Shui J,Cai W,Tang S

    更新日期:2020-12-01 00:00:00

  • Molecular studies on bromovirus capsid protein. VII. Selective packaging on BMV RNA4 by specific N-terminal arginine residuals.

    abstract::An arginine-rich RNA-binding motif (ARM) found at the N-proximal region of Brome mosaic virus (BMV) coat protein (CP) adopts alpha-helical conformation and shares homology with CPs of plant and insect RNA viruses, HIV-Rev and Tat proteins, bacterial antiterminators, and ribosomal splicing factors. The ARM of BMV CP, c...


    pub_type: 杂志文章


    authors: Choi YG,Rao AL

    更新日期:2000-09-15 00:00:00

  • Limitation of Tat-associated transcriptional processivity in HIV-infected PBMC.

    abstract::The ability of HIV to match levels of viral mRNA to the activation state of the host cell may play a role in its ability to persist as well as to replicate. This linkage depends on the function of the viral transcriptional regulatory protein, Tat, which increases the efficiency of RNA elongation (transcriptional proce...


    pub_type: 杂志文章


    authors: Adams M,Wong C,Wang D,Romeo J

    更新日期:1999-05-10 00:00:00

  • Viral RNA load and histological changes in tissues following experimental infection with an arterivirus of possums (wobbly possum disease virus).

    abstract::Tissues from Australian brushtail possums (Trichosurus vulpecula) that had been experimentally infected with wobbly possum disease (WPD) virus (WPDV) were examined to elucidate pathogenesis of WPDV infection. Mononuclear inflammatory cell infiltrates were present in livers, kidneys, salivary glands and brains of WPD-a...


    pub_type: 杂志文章


    authors: Giles J,Perrott M,Roe W,Shrestha K,Aberdein D,Morel P,Dunowska M

    更新日期:2018-09-01 00:00:00

  • Classical swine H1N1 influenza viruses confer cross protection from swine-origin 2009 pandemic H1N1 influenza virus infection in mice and ferrets.

    abstract::The hemagglutinin of the 2009 pandemic H1N1 influenza virus is a derivative of and is antigenically related to classical swine but not to seasonal human H1N1 viruses. We compared the A/California/7/2009 (CA/7/09) virus recommended by the WHO as the reference virus for vaccine development, with two classical swine infl...


    pub_type: 杂志文章


    authors: Min JY,Chen GL,Santos C,Lamirande EW,Matsuoka Y,Subbarao K

    更新日期:2010-12-05 00:00:00

  • Gammaherpesvirus targets peritoneal B-1 B cells for long-term latency.

    abstract::Gammaherpesviruses establish life-long infection in most adults and are associated with the development of B cell lymphomas. While the interaction between gammaherpesviruses and splenic B cells has been explored, very little is known about gammaherpesvirus infection of B-1 B cells, innate-like B cells that primarily r...


    pub_type: 杂志文章


    authors: Rekow MM,Darrah EJ,Mboko WP,Lange PT,Tarakanova VL

    更新日期:2016-05-01 00:00:00

  • Histamine enhances HIV-1-induced modulation of dendritic cells to skew naïve T cell differentiation toward regulatory T cells.

    abstract::Altered cytokine profiles and imbalanced frequencies of CD4(+) T helper cell subsets are thought to be linked with HIV-1/AIDS pathogenesis, but the causes need to be further clarified. Histamine, a biogenic amine with many functions, shows enhancement in HIV-1 infected individuals, which are considered to link with di...


    pub_type: 杂志文章


    authors: Zhai RR,Jiang AP,Wang HB,Ma L,Ren XX,Jiang JF,Wu L,Wei JF,Wang JH

    更新日期:2013-08-01 00:00:00

  • Isolation of mutations of the phage Mu ner gene.

    abstract::A method was devised which allows the easy detection of mutations within the ner gene of Mu DNA. This method is based upon the observation that a transcriptional gene A-galK fusion containing the complete ner gene and the cts62 allele does not express the galK gene in an Escherichia coli strain lacking functional inte...


    pub_type: 杂志文章


    authors: König U,Kirschner P,Schumann W

    更新日期:1988-05-01 00:00:00

  • Phylogenetic analysis of the N8 neuraminidase gene of influenza A viruses.

    abstract::Phylogenetic analysis of the N8 neuraminidase (NA) genes from 18 influenza A viruses, representing equine and avian hosts in different geographic locations, revealed three major lineages: (i) currently circulating equine 2 viruses; (ii) avian viruses isolated in the Eurasian region, including A/Equine/Jilin/1/89, a re...


    pub_type: 杂志文章


    authors: Saito T,Kawaoka Y,Webster RG

    更新日期:1993-04-01 00:00:00

  • Friend murine leukemia virus and spleen focus-forming virus expression in highly malignant interferon-sensitive and interferon-resistant Friend leukemia cells.

    abstract::Analysis of expression of the Friend murine leukemia virus (F-MuLV) and of the spleen focus forming virus (SFFV) has been undertaken in highly malignant interferon (IFN)-sensitive (745) and IFN-resistant (3Cl-8) Friend leukemia cells (FLC), serially passaged intraperitoneally in DBA/2 mice. In vivo passaged 745 cells,...


    pub_type: 杂志文章


    authors: Oppi C,Fiorucci G,Ferrantini M,Battistini A,Belardelli F

    更新日期:1986-04-30 00:00:00

  • Selective inhibition of formation of suppressor glutamine tRNA in Moloney murine leukemia virus-infected NIH-3T3 cells by Avarol.

    abstract::Avarol is a sesquiterpenoid hydroquinone, which displays no inhibitory potencies on mammalian DNA polymerases alpha, beta, and gamma, on mammalian RNA polymerases I, II, and III, or on reverse transcriptases from Moloney murine leukemia virus (Mo-MuLV) and from HIV. For a further elucidation of the antiviral effect of...


    pub_type: 杂志文章


    authors: Kuchino Y,Nishimura S,Schröder HC,Rottmann M,Müller WE

    更新日期:1988-08-01 00:00:00

  • Inhibition of uncoating of tobacco mosaic virus particles in protoplasts from transgenic tobacco plants that express the viral coat protein gene.

    abstract::The uncoating of tobacco mosaic virus (TMV) particles in protoplasts isolated from leaves of transgenic tobacco plants that express the TMV coat protein gene was investigated. Extracts of these protoplasts collected up to 1 hr after inoculation with TMV contained fewer of the complexes ("striposomes") thought to be in...


    pub_type: 杂志文章


    authors: Wu XJ,Beachy RN,Wilson TM,Shaw JG

    更新日期:1990-12-01 00:00:00

  • Specific functions of the Rep and Rep׳ proteins of porcine circovirus during copy-release and rolling-circle DNA replication.

    abstract::The roles of two porcine circovirus replication initiator proteins, Rep and Rep׳, in generating copy-release and rolling-circle DNA replication intermediates were determined. Rep uses the supercoiled closed-circular genome (ccc) to initiate leading-strand synthesis (identical to copy-release replication) and generates...


    pub_type: 杂志文章


    authors: Cheung AK

    更新日期:2015-07-01 00:00:00

  • Phospholipid molecular species profiles of tectiviruses infecting Gram-negative and Gram-positive hosts.

    abstract::The phospholipid (PL) molecular species compositions of bacteriophages PRD1 and Bam35 as well as their respective hosts were determined quantitatively using liquid chromatography/electrospray ionization mass-spectrometry (LC-ESI-MS) and backed up by gas-chromatographic/mass-spectrometric (GC-MS) analysis of the total ...


    pub_type: 杂志文章


    authors: Laurinavicius S,Käkelä R,Somerharju P,Bamford DH

    更新日期:2004-05-01 00:00:00

  • Differential virus restriction patterns of rhesus macaque and human APOBEC3A: implications for lentivirus evolution.

    abstract::The human apolipoprotein B mRNA editing enzyme catalytic peptide-like 3 (APOBEC3; A3) family of proteins (A3A-H) are known to restrict various retroviruses and retroelements, but the full complement of rhesus macaque A3 proteins remains unclear. We report the isolation and characterization of the hA3A homologue from r...


    pub_type: 杂志文章


    authors: Schmitt K,Guo K,Algaier M,Ruiz A,Cheng F,Qiu J,Wissing S,Santiago ML,Stephens EB

    更新日期:2011-10-10 00:00:00

  • Domain structure, intracellular trafficking, and beta2-microglobulin binding of a major histocompatibility complex class I homolog encoded by molluscum contagiosum virus.

    abstract::The MC80R gene of molluscum contagiosum virus (MCV) type 1 encodes a major histocompatibility complex (MHC) class I homolog that lacks several amino-acid residues critical for peptide binding by MHC molecules, contains an unusually long N-terminal hydrophobic domain possibly derived by triplication of a signal peptide...


    pub_type: 杂志文章


    authors: Senkevich TG,Moss B

    更新日期:1998-10-25 00:00:00