Regular ArticleOverexpression and Rapid Purification of Biologically Active Yeast Proliferating Cell Nuclear Antigen
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Genetic variations in the DNA replication origins of human papillomavirus family correlate with their oncogenic potential
2018, Biochimica et Biophysica Acta - General SubjectsCitation Excerpt :The cells were harvested by centrifugation and resuspended in buffer A containing 1 μg/ml of each protease inhibitor including pepstatin, leupeptin, chymostatin, and aprotinin. Induced cells were extracted with the addition of lysozyme following standard protocol [36]. The extract was loaded onto cOmplete His-tag purification resin (Roche, Pleasanton, CA), equilibrated with buffer B.
The Saccharomyces cerevisiae Mlh1-Mlh3 heterodimer is an endonuclease that preferentially binds to holliday junctions
2014, Journal of Biological ChemistryCitation Excerpt :Recombinant Exo1 (D173A) was prepared as described previously (40). PCNA and RFC were expressed and purified from E. coli by minor modifications of previously established procedures (41, 42). We thank Robert Bambara (University of Rochester) and Manju Hingorani (Wesleyan University) for the expression plasmids.
Modulation of DNA synthesis in Saccharomyces cerevisiae nuclear extract by DNA polymerases and the origin recognition complex
2005, Journal of Biological ChemistryCitation Excerpt :Radiolabeled nucleotides ([α-32P]dATP, [α-32P]CTP, and [α-32P]GTP) were obtained from PerkinElmer Life Sciences (Boston, MA). Yeast RPA and recombinant yeast PCNA were purified as described previously (44, 45). Zymolase was purchased from Seikagaku America Inc. (Falmouth, MA).
Biochemical defects in retina-specific human ATP binding cassette transporter nucleotide binding domain 1 mutants associated with macular degeneration
2002, Journal of Biological ChemistryCitation Excerpt :The absence of fortuitous mutations was confirmed by DNA sequencing carried out at the Nucleic Acid Core Facility of Thomas Jefferson University. The resulting recombinant plasmid (pET29aNBD1) was used for expression of the wtNBD1 protein in Escherichia coliBL21(DE3) cells, following a procedure described earlier (32). This plasmid was also used as the parent of all the mutant clones of NBD1.
Sequence-Dependent Interaction of the Human Papillomavirus E2 Protein with the DNA Elements on Its DNA Replication Origin
2023, International Journal of Molecular Sciences