SCF-Mediated Degradation of Claspin Regulates Recovery from the DNA Replication Checkpoint Response
نویسندگان
چکیده
Supplemental Experimental Procedures Plasmids The N-terminal and C-terminal fragments of Claspin were separately amplified by RT-PCR using a cDNA library generated from HEK293T cells. The two RT-PCR products were sequentially inserted into pcDNA3.1-HA tag to obtain a plasmid containing full-length Claspin. Claspin mutants were generated using the QuickChange Site-directed Mutagenesis kit (Stratagene). For retrovirus production, both wild-type Claspin and Claspin mutants were subcloned into the retroviral vector LZRSpBMN by PCR. All cDNAs were sequenced. Plk1 constructs (wild type, K82R and T210D) were kindly provided by Prasad Jallepalli (MSKCC).
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