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  1. Article ; Online: Glioma-amplified sequence KUB3 influences double-strand break repair after ionizing radiation.

    Fischer, Ulrike / Rheinheimer, Stefanie / Krempler, Andrea / Löbrich, Markus / Meese, Eckart

    International journal of oncology

    2013  Volume 43, Issue 1, Page(s) 50–56

    Abstract: Human glioblastomas are characterized by frequent DNA amplifications most often at chromosome regions 7p11.2 and 12q13-15. Although amplification is a well-known hallmark of glioblastoma genetics the function of most amplified genes in glioblastoma ... ...

    Abstract Human glioblastomas are characterized by frequent DNA amplifications most often at chromosome regions 7p11.2 and 12q13-15. Although amplification is a well-known hallmark of glioblastoma genetics the function of most amplified genes in glioblastoma biology is not understood. Previously, we cloned Ku70-binding protein 3 (KUB3) from the amplified domain at 12q13-15. Here, we report that glioblastoma cell cultures with endogenous KUB3 gene amplification and with elevated KUB3 protein expression show an efficient double-strand break (DSB) repair after being irradiated with 1 Gy. A significantly less efficient DSB repair was found in glioma cell cultures without KUB3 amplification and expression. Furthermore, we found that a siRNA-mediated reduction of the endogenous KUB3 expression in glioblastoma cells resulted in a reduction of the repair efficiency. HeLa cells transfected with KUB3 showed an increased DSB repair in comparison to untreated HeLa cells. In addition, KUB3 seems to influence DSB efficiency via the DNA-PK-dependent repair pathway as shown by simultaneous inhibition of KUB3 and DNA-PK. The data provide the first evidence for a link between the level of KUB3 amplification and expression in glioma and DSB repair efficiency.
    MeSH term(s) Antigens, Nuclear/metabolism ; Carrier Proteins/genetics ; DNA Breaks, Double-Stranded/radiation effects ; DNA Repair/radiation effects ; DNA-Binding Proteins/metabolism ; Gene Amplification/radiation effects ; Gene Expression Regulation, Neoplastic/radiation effects ; Glioma/genetics ; Glioma/metabolism ; Glioma/pathology ; HeLa Cells ; Humans ; Ku Autoantigen ; Protein Binding/radiation effects ; RNA, Small Interfering ; Radiation, Ionizing
    Chemical Substances ATP23 protein, human ; Antigens, Nuclear ; Carrier Proteins ; DNA-Binding Proteins ; RNA, Small Interfering ; Xrcc6 protein, human (EC 3.6.4.12) ; Ku Autoantigen (EC 4.2.99.-)
    Language English
    Publishing date 2013-05-13
    Publishing country Greece
    Document type Journal Article
    ZDB-ID 1154403-x
    ISSN 1791-2423 ; 1019-6439
    ISSN (online) 1791-2423
    ISSN 1019-6439
    DOI 10.3892/ijo.2013.1937
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Book ; Online ; Thesis: Analyse der RyREF2-Bindungsstelle im Promotor des porcinen RyR1-Gens und Charakterisierung des porcinen FHL1-Gens

    Krempler, Andrea

    2000  

    Author's details vorgelegt von Andrea Krempler
    Language German
    Size Online-Ressource
    Edition [Elektronische Ressource]
    Document type Book ; Online ; Thesis
    Thesis / German Habilitation thesis Univ., Diss--Göttingen, 2000
    Note Dateiformat: zip, Dateien in unterschiedlichen Formaten
    Database Former special subject collection: coastal and deep sea fishing

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  3. Book ; Online ; Thesis: Anaylse der RyREF2-Bindungsstelle im Promotor des procinen RyR1 Gens und Characterisierung des procinen FHL1-Gens

    Krempler, Andrea

    2000  

    Abstract: Gewebsspezifische Genexpression wird zum größten Teil durch das Zusammenspiel einer Reihe von Transkriptionsfaktoren bewerkstelligt. Um die transkriptionelle Kontrolle muskelspezifischer Genexpression näher zu untersuchen, wurde ein Promoterfragment des ... ...

    Author's details Andrea Krempler
    Abstract Gewebsspezifische Genexpression wird zum größten Teil durch das Zusammenspiel einer Reihe von Transkriptionsfaktoren bewerkstelligt. Um die transkriptionelle Kontrolle muskelspezifischer Genexpression näher zu untersuchen, wurde ein Promoterfragment des procinen Skelettmuskel-Ryanodinrezeptors 1 analysiert (RyR1). Für dieses Promotorfragment konnte in früheren Untersuchungen Bindungsaktivität eines unbekannten Transkriptionsfaktors nachgewiesen werden. Die Anwendung molekularbiologischer und biochemischer Isolationsmethoden zeigte, daß die beschriebene Bindungsaktivität in Gesamtzellextrakten auf Interaktion eines unspezifischen Proteins beruht. Die Analyse von Skelettmuskel-Kernextrakten ergab keine spezifische Bindung eines Faktors im Retentionsgel ...
    Language German
    Size Online-Ressource
    Edition [Electronic ed.]
    Publisher Niedersächsische Staats- und Universitätsbibliothek
    Publishing place Göttingen
    Document type Book ; Online ; Thesis
    Thesis / German Habilitation thesis Univ., Diss.--Göttingen, 2000
    Database Former special subject collection: coastal and deep sea fishing

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  4. Book ; Online ; Thesis: Analyse der RyREF2-Bindungsstelle im Promotor des porcinen RyR1-Gens und Charakterisierung des porcinen FHL1-Gens

    Krempler, Andrea [Verfasser]

    2000  

    Author's details vorgelegt von Andrea Krempler
    Keywords Medizin, Gesundheit ; Medicine, Health
    Subject code sg610
    Document type Book ; Online ; Thesis
    Database Digital theses on the web

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  5. Article ; Online: Elevated radiation-induced γH2AX foci in G2 phase heterozygous BRCA2 fibroblasts.

    Beucher, Andrea / Deckbar, Dorothee / Schumann, Eik / Krempler, Andrea / Frankenberg-Schwager, Marlis / Löbrich, Markus

    Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology

    2011  Volume 101, Issue 1, Page(s) 46–50

    Abstract: Background and purpose: About 5-10% of all breast cancer cases are associated with heterozygous germ-line mutations in the genes encoding BRCA1 and BRCA2. Carriers of such mutations are highly predisposed for developing breast or ovarian cancer and, ... ...

    Abstract Background and purpose: About 5-10% of all breast cancer cases are associated with heterozygous germ-line mutations in the genes encoding BRCA1 and BRCA2. Carriers of such mutations are highly predisposed for developing breast or ovarian cancer and, thus, are advised to undergo regular radio-diagnostic examinations. BRCA1 and BRCA2 are involved in multiple cellular processes including the repair of ionizing radiation (IR)-induced DNA double-strand breaks (DSBs) and different studies addressing the DSB repair capacity of BRCA1+/- or BRCA2+/- cells led to contradictory results.
    Materials and methods: Using the sensitive method of γH2AX foci analysis in combination with cell cycle markers, we specifically measured DSB repair in confluent G0 as well as in exponentially growing G1 and G2 phase primary WT, BRCA1+/- and BRCA2+/- fibroblasts.
    Results: Both BRCA1+/- and BRCA2+/- cells displayed normal DSB repair in G0 and in G1. In contrast, in G2, BRCA2+/- but not BRCA1+/- cells exhibited a decreased DSB repair capacity which was in between that of WT and that of a hypomorphic BRCA2-/- cell line.
    Conclusions: The residual amount of normal BRCA1 seems to be sufficient for efficient DSB repair in all cell cycle phases, while the decreased DSB repair capacity of heterozygous BRCA2 mutations suggests gene dosage effects in G2.
    MeSH term(s) BRCA2 Protein/genetics ; Breast Neoplasms/genetics ; Breast Neoplasms/pathology ; Breast Neoplasms/radiotherapy ; Cell Cycle Checkpoints/genetics ; Cell Cycle Checkpoints/radiation effects ; Cell Line, Tumor/radiation effects ; DNA Breaks, Double-Stranded ; DNA Repair/genetics ; Female ; Fibroblasts/pathology ; Fibroblasts/radiation effects ; G2 Phase/radiation effects ; Heterozygote ; Histones/analysis ; Histones/radiation effects ; Humans ; Radiation Dosage ; Radiation Tolerance ; Radiation, Ionizing
    Chemical Substances BRCA2 Protein ; BRCA2 protein, human ; H2AX protein, human ; Histones
    Language English
    Publishing date 2011-06-12
    Publishing country Ireland
    Document type Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 605646-5
    ISSN 1879-0887 ; 0167-8140
    ISSN (online) 1879-0887
    ISSN 0167-8140
    DOI 10.1016/j.radonc.2011.05.043
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: An imperfect G2M checkpoint contributes to chromosome instability following irradiation of S and G2 phase cells.

    Krempler, Andrea / Deckbar, Dorothee / Jeggo, Penny A / Löbrich, Markus

    Cell cycle (Georgetown, Tex.)

    2007  Volume 6, Issue 14, Page(s) 1682–1686

    Abstract: DNA double strand break (DSB) repair and checkpoint control represent two major mechanisms that function to reduce chromosomal instability following ionizing irradiation (IR). Ataxia telangiectasia (A-T) cells have long been known to have defective ... ...

    Abstract DNA double strand break (DSB) repair and checkpoint control represent two major mechanisms that function to reduce chromosomal instability following ionizing irradiation (IR). Ataxia telangiectasia (A-T) cells have long been known to have defective checkpoint responses. Recent studies have shown that they also have a DSB repair defect following IR raising the issue of how ATM's repair and checkpoint functions interplay to maintain chromosomal stability. A-T and Artemis cells manifest an identical and epistatic repair defect throughout the cell cycle demonstrating that ATM's major repair defect following IR represents Artemis-dependent end-processing. Artemis cells show efficient G(2)/M checkpoint induction and a prolonged arrest relative to normal cells. Following irradiation of G(2) cells, this checkpoint is dependent on ATM and A-T cells fail to show checkpoint arrest. In contrast, cells irradiated during S phase initiate a G(2)/M checkpoint which is independent of ATM and, significantly, both Artemis and A-T cells show a prolonged arrest at the G(2)/M checkpoint likely reflecting their repair defect. Strikingly, the G(2)/M checkpoint is released before the completion of repair when approximately 10-20 DSBs remain both for S phase and G(2) phase irradiated cells. This defined sensitivity level of the G(2)/M checkpoint explains the prolonged arrest in repair-deficient relative to normal cells and provides a conceptual framework for the cooperative phenotype between checkpoint and repair functions in maintaining chromosomal stability.
    MeSH term(s) Ataxia Telangiectasia Mutated Proteins ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/metabolism ; Cell Division/physiology ; Chromosomal Instability ; DNA/genetics ; DNA/metabolism ; DNA/radiation effects ; DNA Damage ; DNA Repair ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/metabolism ; Endonucleases ; G2 Phase/physiology ; Humans ; Nuclear Proteins/genetics ; Nuclear Proteins/metabolism ; Protein Serine-Threonine Kinases/genetics ; Protein Serine-Threonine Kinases/metabolism ; Radiation, Ionizing ; S Phase/physiology ; Tumor Suppressor Proteins/genetics ; Tumor Suppressor Proteins/metabolism
    Chemical Substances Cell Cycle Proteins ; DNA-Binding Proteins ; Nuclear Proteins ; Tumor Suppressor Proteins ; DNA (9007-49-2) ; ATM protein, human (EC 2.7.11.1) ; ATR protein, human (EC 2.7.11.1) ; Ataxia Telangiectasia Mutated Proteins (EC 2.7.11.1) ; Protein Serine-Threonine Kinases (EC 2.7.11.1) ; DCLRE1C protein, human (EC 3.1.-) ; Endonucleases (EC 3.1.-)
    Language English
    Publishing date 2007-05-21
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2146183-1
    ISSN 1551-4005 ; 1538-4101 ; 1554-8627
    ISSN (online) 1551-4005
    ISSN 1538-4101 ; 1554-8627
    DOI 10.4161/cc.6.14.4480
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Zs.A 5881: Show issues Location:
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  7. Article: Radiation-induced HPRT mutations resulting from misrejoined DNA double-strand breaks.

    Rothkamm, Kai / Gunasekara, Kut / Warda, Salam A / Krempler, Andrea / Löbrich, Markus

    Radiation research

    2008  Volume 169, Issue 6, Page(s) 639–648

    Abstract: DNA double-strand breaks (DSBs) are the most severe lesions induced by ionizing radiation, and unrejoined or misrejoined DSBs can lead to cell lethality, mutations and the initiation of tumorigenesis. We have investigated X-ray- and alpha-particle- ... ...

    Abstract DNA double-strand breaks (DSBs) are the most severe lesions induced by ionizing radiation, and unrejoined or misrejoined DSBs can lead to cell lethality, mutations and the initiation of tumorigenesis. We have investigated X-ray- and alpha-particle-induced mutations that inactivate the hypoxanthine guanine phosphoribosyltransferase (HPRT) gene in human bladder carcinoma cells and in hTERT-immortalized human fibroblasts. Fifty to 80% of the mutants analyzed exhibited partial or total deletions of the 9 exons of the HPRT locus. The remaining mutants retained unaltered PCR products of all 9 exons but often displayed a failure to amplify the HPRT cDNA. Hybridization analysis of a 2-Mbp NotI fragment spanning the HPRT gene with a probe 200 kbp distal to the HPRT locus indicated altered fragment sizes in most of the mutants with a wild-type PCR pattern. These mutants likely contain breakpoints for genomic rearrangements in the intronic sequences of the HPRT gene that allow the amplification of the exons but prevent HPRT cDNA amplification. Additionally, mutants exhibiting partial and total deletions of the HPRT exons also frequently displayed altered NotI fragments. Interestingly, all mutations were very rarely associated with interchromosomal exchanges analyzed by FISH. Collectively, our data suggest that intrachromosomal genomic rearrangements on the Mbp scale represent the prevailing type of radiation-induced HPRT mutations.
    MeSH term(s) Alpha Particles ; Cell Line, Tumor ; Chromosome Mapping ; DNA Breaks, Double-Stranded ; DNA Primers/chemistry ; DNA Repair ; DNA, Complementary/metabolism ; Dose-Response Relationship, Radiation ; Fibroblasts/metabolism ; Humans ; Hypoxanthine Phosphoribosyltransferase/genetics ; In Situ Hybridization, Fluorescence ; Mutation ; Nucleic Acid Hybridization ; X-Rays
    Chemical Substances DNA Primers ; DNA, Complementary ; Hypoxanthine Phosphoribosyltransferase (EC 2.4.2.8)
    Language English
    Publishing date 2008-06
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80322-4
    ISSN 1938-5404 ; 0033-7587
    ISSN (online) 1938-5404
    ISSN 0033-7587
    DOI 10.1667/RR1185.1
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article: Targeted deletion of the Tsg101 gene results in cell cycle arrest at G1/S and p53-independent cell death.

    Krempler, Andrea / Henry, MaLinda D / Triplett, Aleata A / Wagner, Kay-Uwe

    The Journal of biological chemistry

    2002  Volume 277, Issue 45, Page(s) 43216–43223

    Abstract: The tumor susceptibility gene 101 (Tsg101) was originally discovered in a screen for potential tumor suppressors using insertional mutagenesis in immortalized fibroblasts. To investigate essential functions of this gene in cell growth and neoplastic ... ...

    Abstract The tumor susceptibility gene 101 (Tsg101) was originally discovered in a screen for potential tumor suppressors using insertional mutagenesis in immortalized fibroblasts. To investigate essential functions of this gene in cell growth and neoplastic transformation, we derived primary mouse embryonic fibroblasts from Tsg101 conditional knockout mice. Expression of Cre recombinase from a retroviral vector efficiently down-regulated Tsg101. The deletion of Tsg101 caused growth arrest and cell death but did not result in increased proliferation and cellular transformation. Inactivation of p53 had no influence on the deleterious phenotype, but Tsg101(-/-) cells were rescued through expression of exogenous Tsg101. Fluorescence-activated cell sorting, proliferation assays, and Western blot analysis of crucial regulators of the cell cycle revealed that Tsg101 deficiency resulted in growth arrest at the G(1)/S transition through inactivation of cyclin-dependent kinase 2. As a consequence, DNA replication was not initiated in Tsg101-deficient cells. Our results clearly demonstrate that Tsg101 is not a primary tumor suppressor in mouse embryonic fibroblasts. However, the protein is crucial for cell proliferation and cell survival.
    MeSH term(s) Animals ; Cell Cycle/physiology ; Cell Death/physiology ; Cell Division ; Cell Survival ; Cloning, Molecular ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/physiology ; Embryo, Mammalian ; Endosomal Sorting Complexes Required for Transport ; Fibroblasts/cytology ; G1 Phase/physiology ; Leucine Zippers ; Mice ; Mutagenesis ; Recombinant Proteins/metabolism ; S Phase/physiology ; Sequence Deletion ; Transcription Factors/genetics ; Transcription Factors/physiology ; Tumor Suppressor Protein p53/metabolism
    Chemical Substances DNA-Binding Proteins ; Endosomal Sorting Complexes Required for Transport ; Recombinant Proteins ; Transcription Factors ; Tsg101 protein ; Tumor Suppressor Protein p53
    Language English
    Publishing date 2002-08-29
    Publishing country United States
    Document type Journal Article ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 2997-x
    ISSN 1083-351X ; 0021-9258
    ISSN (online) 1083-351X
    ISSN 0021-9258
    DOI 10.1074/jbc.M207662200
    Database MEDical Literature Analysis and Retrieval System OnLINE

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    Uc I Zs.108: Show issues Location:
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  9. Article: Chromosome breakage after G2 checkpoint release.

    Deckbar, Dorothee / Birraux, Julie / Krempler, Andrea / Tchouandong, Leopoldine / Beucher, Andrea / Walker, Sarah / Stiff, Tom / Jeggo, Penny / Löbrich, Markus

    The Journal of cell biology

    2007  Volume 176, Issue 6, Page(s) 749–755

    Abstract: DNA double-strand break (DSB) repair and checkpoint control represent distinct mechanisms to reduce chromosomal instability. Ataxia telangiectasia (A-T) cells have checkpoint arrest and DSB repair defects. We examine the efficiency and interplay of ATM's ...

    Abstract DNA double-strand break (DSB) repair and checkpoint control represent distinct mechanisms to reduce chromosomal instability. Ataxia telangiectasia (A-T) cells have checkpoint arrest and DSB repair defects. We examine the efficiency and interplay of ATM's G2 checkpoint and repair functions. Artemis cells manifest a repair defect identical and epistatic to A-T but show proficient checkpoint responses. Only a few G2 cells enter mitosis within 4 h after irradiation with 1 Gy but manifest multiple chromosome breaks. Most checkpoint-proficient cells arrest at the G2/M checkpoint, with the length of arrest being dependent on the repair capacity. Strikingly, cells released from checkpoint arrest display one to two chromosome breaks. This represents a major contribution to chromosome breakage. The presence of chromosome breaks in cells released from checkpoint arrest suggests that release occurs before the completion of DSB repair. Strikingly, we show that checkpoint release occurs at a point when approximately three to four premature chromosome condensation breaks and approximately 20 gammaH2AX foci remain.
    MeSH term(s) Ataxia Telangiectasia Mutated Proteins ; Cell Cycle Proteins/genetics ; Cell Cycle Proteins/physiology ; Cell Line ; Chromosome Breakage ; DNA Breaks, Double-Stranded ; DNA Repair ; DNA-Binding Proteins/genetics ; DNA-Binding Proteins/physiology ; Endonucleases ; G2 Phase/physiology ; Humans ; Nuclear Proteins/physiology ; Protein Serine-Threonine Kinases/genetics ; Protein Serine-Threonine Kinases/physiology ; Tumor Suppressor Proteins/genetics ; Tumor Suppressor Proteins/physiology
    Chemical Substances Cell Cycle Proteins ; DNA-Binding Proteins ; Nuclear Proteins ; Tumor Suppressor Proteins ; ATM protein, human (EC 2.7.11.1) ; Ataxia Telangiectasia Mutated Proteins (EC 2.7.11.1) ; Protein Serine-Threonine Kinases (EC 2.7.11.1) ; DCLRE1C protein, human (EC 3.1.-) ; Endonucleases (EC 3.1.-)
    Language English
    Publishing date 2007-03-12
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 218154-x
    ISSN 1540-8140 ; 0021-9525
    ISSN (online) 1540-8140
    ISSN 0021-9525
    DOI 10.1083/jcb.200612047
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article: Generation of a conditional knockout allele for the Janus kinase 2 (Jak2) gene in mice.

    Krempler, Andrea / Qi, Yongyue / Triplett, Aleata A / Zhu, Jianqiong / Rui, Hallgeir / Wagner, Kay-Uwe

    Genesis (New York, N.Y. : 2000)

    2004  Volume 40, Issue 1, Page(s) 52–57

    Abstract: To study biologically relevant functions of the Janus kinase 2 (Jak2) in multiple cytokine and hormone receptor signal transduction pathways, we generated a conditional knockout (floxed) allele of this gene by placing loxP sites around the first coding ... ...

    Abstract To study biologically relevant functions of the Janus kinase 2 (Jak2) in multiple cytokine and hormone receptor signal transduction pathways, we generated a conditional knockout (floxed) allele of this gene by placing loxP sites around the first coding exon of Jak2. Homozygous floxed animals developed normally and exhibited no phenotypic abnormalities. The conversion of the floxed allele into a null mutation was achieved by transmitting the targeted allele through the female germline of MMTV-Cre (line A) mice. Embryos that carry two Jak2 null alleles died around midgestation and exhibited impaired definitive erythropoiesis, which is a hallmark of Jak2 deficiency reported previously in conventional knockouts. This observation suggested that the Cre-mediated deletion of the first coding exon results in a true null mutation that is incapable of mediating signals through the erythropoietin receptor. Using mouse embryonic fibroblasts derived from Jak2 null embryos and their wildtype littermate controls, we demonstrated that Jak2-deficiency decouples growth hormone-receptor signaling from its downstream mediators, the signal transducer and activator of transcription (Stat) 5a and 5b.
    MeSH term(s) Animals ; Erythropoiesis/genetics ; Exons/genetics ; Female ; Fetal Death/genetics ; Gene Deletion ; Janus Kinase 2 ; Mammary Tumor Virus, Mouse/genetics ; Mice ; Mice, Knockout ; Pregnancy ; Protein-Tyrosine Kinases/deficiency ; Protein-Tyrosine Kinases/genetics ; Proto-Oncogene Proteins/deficiency ; Proto-Oncogene Proteins/genetics
    Chemical Substances Proto-Oncogene Proteins ; Protein-Tyrosine Kinases (EC 2.7.10.1) ; Jak2 protein, mouse (EC 2.7.10.2) ; Janus Kinase 2 (EC 2.7.10.2)
    Language English
    Publishing date 2004-09
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 2004544-X
    ISSN 1526-968X ; 1526-954X
    ISSN (online) 1526-968X
    ISSN 1526-954X
    DOI 10.1002/gene.20063
    Database MEDical Literature Analysis and Retrieval System OnLINE

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