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Checkpoint kinase 2 is required for efficient immunoglobulin diversification.

Maintenance of genome integrity relies on multiple DNA repair pathways as well as on checkpoint regulation. Activation of the checkpoint kinases Chk1 and Chk2 by DNA damage triggers cell cycle arrest and improved DNA repair, or apoptosis in case of excessive damage. Chk1 and Chk2 have been reported to act in a complementary or redundant fashion, depending on the physiological context. During secondary immunoglobulin (Ig) diversification in B lymphocytes, DNA damage is abundantly introduced by activation-induced cytidine deaminase (AID) and processed to mutations in a locus-specific manner by several error-prone DNA repair pathways. We have previously shown that Chk1 negatively regulates Ig somatic hypermutation by promoting error-free homologous recombination and Ig gene conversion. We now report that Chk2 shows opposite effects to Chk1 in the regulation of these processes. Chk2 inactivation in B cells leads to decreased Ig hypermutation and Ig class switching, and increased Ig gene conversion activity. This is linked to defects in non-homologous end joining and increased Chk1 activation upon interference with Chk2 function. Intriguingly, in the context of physiological introduction of substantial DNA damage into the genome during Ig diversification, the 2 checkpoint kinases thus function in an opposing manner, rather than redundantly or cooperatively.
Kathrin Davari, Samantha Frankenberger, Angelika Schmidt, Nils-Sebastian Tomi, Berit Jungnickel

1720 related Products with: Checkpoint kinase 2 is required for efficient immunoglobulin diversification.

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Simplified PCR set-up using a frozen preformulated mix for the detection of cytomegalovirus.

A simple PCR set-up for the detection of cytomegalovirus in clinical specimen was developed. All components of the PCR master mix including Taq DNA polymerase, uracil N-glycosilase, and primers were preformulated and stored frozen in aliquots. After thawing the master mix aliquots, the PCR was immediately started after the addition of sample DNA. This method gave excellent reproducible PCR-results without loss of enzyme activities following storage at -20 degrees C for at least 4 months.
B Kofler, A Klausegger

2063 related Products with: Simplified PCR set-up using a frozen preformulated mix for the detection of cytomegalovirus.