Cell cycle dependent expression of Plk1 in synchronized porcine
fetal fibroblasts

J 2003

Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts

DVOŘÁK, Petr, M. ANGER, W. A. KUES, J. KLÍMA, M. MIELENZ et. al.

Základní údaje

Originální název

Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts

Autoři

DVOŘÁK, Petr, M. ANGER, W. A. KUES, J. KLÍMA, M. MIELENZ, M. KUBELKA, J. MOTLÍK, M. EŠNER, J. W. CARNWATH a H. NIEMANN

Vydání

Molecular reproduction and development, 2003, 1040-452X

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

Genetika a molekulární biologie

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 2.543

Organizační jednotka

Lékařská fakulta

Klíčová slova anglicky

cell

Štítky

cell

Změněno: 1. 2. 2006 13:45, prof. Ing. Petr Dvořák, CSc.

Anotace

V originále

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.

Citovat

DVOŘÁK, Petr, M. ANGER, W. A. KUES, J. KLÍMA, M. MIELENZ, M. KUBELKA, J. MOTLÍK, M. EŠNER, J. W. CARNWATH a H. NIEMANN. Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts. Molecular reproduction and development. 2003, roč. 65, č. 3, s. 245-253. ISSN 1040-452X.

@article{618184,
   author = {Dvořák, Petr and Anger, M. and Kues, W. A. and Klíma, J. and Mielenz, M. and Kubelka, M. and Motlík, J. and Ešner, M. and Carnwath, J. W. and Niemann, H.},
   article_number = {3},
   keywords = {cell},
   language = {eng},
   issn = {1040-452X},
   journal = {Molecular reproduction and development},
   title = {Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts},
   volume = {65},
   year = {2003}
}

TY  - JOUR
ID  - 618184
AU  - Dvořák, Petr - Anger, M. - Kues, W. A. - Klíma, J. - Mielenz, M. - Kubelka, M. - Motlík, J. - Ešner, M. - Carnwath, J. W. - Niemann, H.
PY  - 2003
TI  - Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts
JF  - Molecular reproduction and development
VL  - 65
IS  - 3
SP  - 245-253
EP  - 245-253
SN  - 1040452X
KW  - cell
N2  - 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.
ER  -

DVOŘÁK, Petr, M. ANGER, W. A. KUES, J. KLÍMA, M. MIELENZ, M. KUBELKA, J. MOTLÍK, M. EŠNER, J. W. CARNWATH a H. NIEMANN. Cell cycle dependent expression of Plk1 in synchronized porcine fetal fibroblasts. \textit{Molecular reproduction and development}. 2003, roč.~65, č.~3, s.~245-253. ISSN~1040-452X.

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