DVOŘÁK, Petr, M. ANGER, W. A. KUES, J. KLÍMA, M. MIELENZ, M. KUBELKA, J. MOTLÍK, M. EŠNER, J. W. CARNWATH and H. NIEMANN. Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts. Molecular reproduction and development. 2003, vol. 65, No 3, p. 245-253. ISSN 1040-452X.
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Basic information
Original name Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts
Authors DVOŘÁK, Petr, M. ANGER, W. A. KUES, J. KLÍMA, M. MIELENZ, M. KUBELKA, J. MOTLÍK, M. EŠNER, J. W. CARNWATH and H. NIEMANN.
Edition Molecular reproduction and development, 2003, 1040-452X.
Other information
Original language English
Type of outcome Article in a journal
Field of Study Genetics and molecular biology
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 2.543
Organization unit Faculty of Medicine
Keywords in English cell
Tags cell
Changed by Changed by: prof. Ing. Petr Dvořák, CSc., učo 47260. Changed: 1/2/2006 13:45.
Abstract
Enzymes of the Polo-like kinase (Plk) family are active in the pathways controlling mitosis in several species. We have cloned cDNA fragments of the porcine homologues of Plk1, Plk2, and Plk3 employing fetal fibroblasts as source. All three partial cDNAs showed high sequence homology with their mouse and human counterparts and contained the Polo box, a domain characteristic for all Polo kinases. The expression levels of Plk1 mRNA at various points of the cell cycle in synchronized porcine fetal fibroblasts were analyzed by both RT-PCR and the ribonuclease protection assay. Plk1 mRNA was barely detectable in G0 and G1, increased during S phase and peaked after the G2/M transition. A monoclonal antibody was generated against an in vitro expressed porcine Plk1-protein fragment and used to detect changes in Plk1 expression at the protein level. Plk1 protein was first detected by immunoblotting at the beginning of S phase and was highest after the G2/M transition. In summary, the Plk1 expression pattern in the pig is similar to that reported for other species. The absence of Plk1 mRNA and protein appears to be a good marker for G0/G1 and thus for the selection of donor cells for nuclear transfer based somatic cloning. Copyright 2003 Wiley-Liss, Inc.
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