Your email was sent successfully. Check your inbox.

An error occurred while sending the email. Please try again.

Proceed reservation?

Export
  • 1
    Keywords: NEUTRALIZING ANTIBODIES ; ESCHERICHIA-COLI ; VIRUS-LIKE PARTICLES ; CYTOTOXIC T-LYMPHOCYTES ; ELISA ; capsomeres ; CONTROLLED-TRIAL ; MAJOR CAPSID PROTEIN ; HPV-16 ; Chloroplast transformation ; TRANSGENIC PLANTS ; CYTOPLASMIC MALE-STERILITY ; L1_ 2xCysM gene ; Male sterility ; NICOTIANA-TABACUM ; Plastids
    Abstract: Certain types of human papillomaviruses (HPV) are causatively associated with cervical carcinoma, the second most common cancer in women worldwide. Due to limitations in the availability of currently used virus-like particle (VLP)-based vaccines against HPV to women of developing countries, where most cases of cervical cancer occur, the development of a cost-effective second-generation vaccine is a necessity. Capsomeres have recently been demonstrated to be highly immunogenic and to have a number of advantages as a potential cost-effective alternative to VLP-based HPV vaccines. We have expressed a mutated HPV-16 L1 (L1_2xCysM) gene that retained the ability to assemble L1 protein to capsomeres in tobacco chloroplasts. The recombinant protein yielded up to 1.5% of total soluble protein. The assembly of capsomeres was examined and verified by cesium chloride density gradient centrifugation and sucrose sedimentation analysis. An antigen capture enzyme-linked immunosorbent assay confirmed the formation of capsomeres by using a conformation-specific monoclonal antibody which recognized the conformational epitopes. Transplastomic tobacco plants exhibited normal growth and morphology, but all such lines showed male sterility in the T(0), T(1) and T(2) generations. Taken together, these results indicate the possibility of producing a low-cost capsomere-based vaccine by plastids.
    Type of Publication: Journal article published
    PubMed ID: 20563641
    Signatur Availability
    BibTip Others were also interested in ...
  • 2
    Keywords: APOPTOSIS ; CANCER ; CELLS ; EXPRESSION ; proliferation ; CELL ; Germany ; human ; INFORMATION ; GENE ; GENOME ; PROTEIN ; PROTEINS ; transcription ; DIFFERENTIATION ; TISSUE ; MICE ; DNA ; TISSUES ; SKIN ; BINDING ; BIOLOGY ; MOLECULAR-BIOLOGY ; papillomavirus ; TARGET ; virus ; NO ; TRANSGENIC MICE ; PROMOTER ; PROMOTERS ; cervical cancer ; CERVICAL-CANCER ; REGION ; human papillomavirus ; HIGH-RISK ; HPV ; transactivation ; ONCOGENE ; HUMAN-PAPILLOMAVIRUS ; CERVICAL-CARCINOMA ; NETHERLANDS ; REPRESSION ; molecular biology ; molecular ; RE ; PATTERN ; LEVEL ; analysis ; methods ; EVENTS ; LOSSES ; UBIQUITIN ; CANCERS ; microbiology ; E2 PROTEIN ; HUMAN PAPILLOMAVIRUSES ; animal model ; host ; viral ; UPSTREAM ; UBIQUITIN-C ; biotechnology ; E2 ; READING FRAMES ; E6 PROMOTER ; PAPILLOMAVIRUS DNA-REPLICATION ; ubiquitin C promoter
    Abstract: The E2 early protein of human papillomaviruses (HPV) has been found associated with the mitotic spindle therefore being implicated in the partition of the replicated viral DNA to daughter cells. In addition, E2 proteins bind to the upstream regulatory region of the virus and to cellular promoters modulating thereby cellular transcription and differentiation. In many cervical cancers, the E2 reading frame is interrupted upon incorporation of the viral genome into the host DNA. This results in the loss of the E2 mediated transcriptional repression and uncontrolled expression of the viral oncogenes. All these results have been obtained in transfected cells but no information is available on the E2 effects in the context of the entire organism. Transgenic mice were generated expressing the E2 protein of HPV11 under the control of the Ubiquitin C promoter. E2 mRNA is present in all mice tissues analysed and the E2 protein expressed in the skin (the target tissue of HPV11) was shown by Western blotting, albeit at a very low level. Analysis of the transgenic mice shows no major histological changes in the skin or all other tissues investigated. These data indicate that in transgenic mice the human papillomavirus type 11 E2 does not grossly modulate cellular proliferation or differentiation events
    Type of Publication: Journal article published
    PubMed ID: 17701441
    Signatur Availability
    BibTip Others were also interested in ...
  • 3
    Keywords: MICE ; INFORMATION ; NETHERLANDS ; MANAGEMENT ; BIOLOGY ; methods
    Type of Publication: Meeting abstract published
    Signatur Availability
    BibTip Others were also interested in ...
  • 4
  • 5
    Keywords: RECEPTOR ; EXPRESSION ; CELL ; Germany ; GENERATION ; SYSTEM ; GENE-EXPRESSION ; RNA ; EFFICIENCY ; MICE ; TRANSCRIPTION FACTOR ; BIOLOGY ; MOLECULAR-BIOLOGY ; ACIDS ; TRANSGENIC MICE ; NUCLEAR-FACTOR ; DESIGN ; VECTOR ; LINE ; DELIVERY ; MAMMALIAN-CELLS ; STEM-CELLS ; NETHERLANDS ; molecular biology ; molecular ; RE ; INTERFERENCE ; RNA INTERFERENCE ; GAMMA ; LENTIVIRAL VECTOR ; methods ; NUCLEAR ; function ; TRANSMISSION ; microbiology ; lentiviral vectors ; MicroRNAs ; mosaicism ; biotechnology ; INTERFERING RNAS ; SIRNAS ; Transgenesis ; TRANSGENIC RATS
    Abstract: We have used a lentiviral delivery system (LentiLox3.7) to generate transgenic mice harbouring RNA interference (RNAi) against the hepatocyte nuclear factor 4 gamma (HNF4(gamma)). HNF4(gamma) is a nuclear receptor with unknown function. Our analyses performed on founder (F-0) and first generation (F-1) mice revealed mosaicism in F-0 founders and a low efficiency of transgenesis (6%) in F-1 mice. These data, together with the observation of multiple silenced transgenes, do not favour the use of LentiLox3.7 lentivirus for transgenesis. Despite the low efficiency of transgenesis, we achieved a tissue-dependent knockdown of HNF4(gamma) expression in some mice
    Type of Publication: Journal article published
    PubMed ID: 17682835
    Signatur Availability
    BibTip Others were also interested in ...
  • 6
    Keywords: CELLS ; CELL ; Germany ; human ; MODEL ; GENE ; validation ; MICE ; DNA ; murine ; recombination ; MOUSE ; CHINESE-HAMSTER CELLS ; MAMMALIAN-CELLS ; KNOCK-IN MICE ; MOUSE MODEL ; EMBRYONIC STEM-CELLS ; HOMOLOGOUS RECOMBINATION ; DOUBLE-STRAND BREAKS ; PARP-1 ; gene targeting ; ES CELLS ; ADP-RIBOSYLTRANSFERASE ; APRT LOCUS ; BACTERIAL ARTIFICIAL CHROMOSOMES ; POLY(ADP-RIBOSE) POLYMERASE-1
    Abstract: Here we report an approach to generate a knock-in mouse model using an 'ends-out' gene replacement vector to substitute the murine Parp-1 (mParp-1) coding sequence (32 kb) with its human orthologous sequence (46 kb). Unexpectedly, examination of mutant ES cell clones and mice revealed that site-specific homologous recombination was mimicked in three independently generated ES cell clones by bidirectional extension of the vector homology arms using the endogenous mParp-1-flanking sequences as templates. This was followed by adjacent integration of the targeting vector, thus leaving the endogenous mParp-1 locus functional. A related phenomenon termed 'ectopic gene targeting' has so far only been described for 'ends-in' integration-type vectors in non-ES cell gene targeting. We provide reliable techniques to detect such ectopic gene targeting which represents an unexpected caveat in mouse genetic engineering that should be considered in the design and validation strategy of future gene knock-in approaches
    Type of Publication: Journal article published
    PubMed ID: 19034683
    Signatur Availability
    BibTip Others were also interested in ...
  • 7
    Keywords: NOMENCLATURE ; GUIDELINES ; cryopreservation ; VERSION ; TRANSGENIC MOUSE EMBRYOS ; RULES
    Abstract: The number of genetically modified mice is increasing rapidly. Several limitations when working with these animals are to be considered: small colonies, the continued danger of loss, often a limited breeding-success, the need to keep those mutants in stock, difficult and costly import-procedures, and also a major (scientific) value of those mutants often available only with major restrictions. To gather relevant information about all active and archived genetically modified mouse lines available in-house (〉1.500) and to deal with a unique resource for several, quite different purposes, a data base was developed enabling optimum knowledge management and easy access. The data base covers also legal restraints and is being linked with the institutional publication repository. To identify the lines available detailed information is provided for each line, as the international designation, a short name, the characterization/description, and the genetic modification including the technique used therefore. The origin of the mutation (gene-ID# and donor organism), the origin of regulatory elements and their donors are listed as well as the genetic background, back-cross generation, phenotype, possible publications, keywords, and some in-house information. Also aspects of animal welfare, obligations to record genetically modified organisms, and technology transfer are displayed; the latter to make licenses possible (if legally permitted). Material transfer agreements, patents, or legal restrictions are listed. This data base helps to avoid double-imports, saves animals and costs since a redundant generation or import can be omitted. However, this is a contribution to the 3R principles developed by Russell and Burch.
    Type of Publication: Journal article published
    PubMed ID: 22302697
    Signatur Availability
    BibTip Others were also interested in ...
  • 8
    ISSN: 1573-9368
    Keywords: transgenic mice ; insertional mutagenesis ; microphthalmia ; depigmentation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic mice were produced by microinjection of a humanAγ-globin gene construct containing site 2 of the locus control region and theAγ-globin gene with its 3′ enhancer sequence. One transgenic mouse line 95′HS2γen91) displayed an altered phenotype when the insertion event of this transgenic line was homozygous. These animals lack the normal pigmentation seen in their hemizygous and non-transgenic littermates, thus appearing white with unpigmented eyes. In addition, their eyes are underdeveloped, consistent with the phenotype associated with mutations at themicrophthalmia (mi) locus. Backcrosses of transgenic mice withmi mutant mice result in phenotypes showing a lack of complementation, demonstrating that the site of transgene insertion is allelic withmi. Electron microscopic analysis of hair follicles and culturing of melanocytes from the skin of transgenic animals reveals an absence of cutaneous melanocytes in homozygotes and aberrant growth and morphology of the melanocytes isolated from hemizygous animals. The results presented here summarize the effects of this new allele of themi locus.
    Type of Medium: Electronic Resource
    Signatur Availability
    BibTip Others were also interested in ...
  • 9
    ISSN: 1573-9368
    Keywords: transposon ; Activator ; Dissociation ; cell-autonomous ; spectinomycin ; tomato ; phosphinothricin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell-autonomous genes have been used to monitor the excision of both endogenous transposons in maize andAntirrhinum, and transposons introduced into transgenic plants. In tobacco andArabidopsis, the streptomycin phosphotransferase (SPT) gene reveals somatic excision of the maize transposonActivator (Ac) as green sectors on a white background in cotyledons of seedlings germinated in the presence of streptomycin. Cotyledons of tomato seedlings germinated on streptomycin-containing medium do not bleach, suggesting that a different assay for transposon excision in tomato is desirable. We have tested the use of the spectinomycin resistance (SPEC) gene (aadA) and a Basta resistance (BAR) gene (phosphinothricin acetyltransferase, or PAT) for monitoring somatic excision ofAc in tobacco and tomato. Both genetic and molecular studies demonstrate that genotypically variegated individuals that carry clones of cells from whichAc orDs have excised from either SPEC or BAR genes, can be phenotypically completely resistant to the corresponding antibiotic. This demonstrates that these genes act non-cell-autonomously, in contrast to the SPT gene in tobacco. Possible reasons for this difference are discussed.
    Type of Medium: Electronic Resource
    Signatur Availability
    BibTip Others were also interested in ...
  • 10
    ISSN: 1573-9368
    Keywords: transgenic ; fetal haemoglobin ; globin ; haemoglobinopathies
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic mice were produced from two 13 kb constructs containing the fetal (γ) globin genes. Each construct consisted of a Gγ globin gene linked to one of two alternative Aγ genes. The first construct contained a normal Gγ gene plus and Aγ gene with the −198T→C hereditary persistence of fetal haemoglobin (HPFH) mutation. In the second, a normal Gγ gene was linked to a Aγ gene with a−222 to −225 four base pair deletion in the promoter. This latter mutation has been associated with low Aγ expression in humans. Both Aγ genes in these constructs also contained the 3′ flanking enhancer. The two different constructs showed expression throughout gestation from day 11 yolk sac, through the fetal period and for a variable time during the first three postnatal weeks. Thereafter, no expression of any of the γ-globin genes was seen in adult erythroid tissues. Transcription from the normal Gγ and HPFH Aγ genes in the first construct paralleled each other in developmental timing, with a proportionate excess of Aγ more evident in later gestation. Gγ and Aγ mRNA transcripts from the normal Gγ+4 bp deletional Aγ construct were unable to be distinguished because of a 3′ Gγ-like conversion in the deletional Aγ gene. Combined γ-globin expression from the two genes in this second construct was detectable until just after birth, as seen with the individual genes in the first construct. Previous work has shown that when individual Gγ and Aγ transgenes are introduced into mice unlinked, without the locus control region (LCR), embryonic expression only is seen. In the present study, γ-globin expression was evident from both constructs throughout the fetal period of erythropoiesis. This suggests that the additional DNA sequences flanking the γ-globin genes included in these larger constructs are capable of supporting fetal recruitment of the γ-globin genes, independently of the LCR.
    Type of Medium: Electronic Resource
    Signatur Availability
    BibTip Others were also interested in ...
Close ⊗
This website uses cookies and the analysis tool Matomo. More information can be found here...