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Isolation and identification of proteins binding to the major breakpoint region(mbr) of bcl2 gene
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This work was supported by grants from the National Natural Science Foundation of China(30500585)and the Natural Science Foundation of Jiangsu Province(BK2008450)

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    Objective: We have previously found that mbr is a regulatory element of the bcl2 gene. The objective of this study is to isolate and identify the proteins binding to the 37 mbr in the 3′-end of the mbr. Methods: Streptavidin magnetic particles were ligated to concatameric oligonucleotides of 37 mbr and incubated with the nuclear extracts of Jurkat cells. The DNA-binding proteins were eluted and then resolved by SDS-PAGE. After silver staining, the protein bands were excised and subjected to MALDI-TOF MS. Results: Several protein bands were detected after the isolation with magnetic particles, and Splicing factor, proline- and glutamine-rich(SFPQ), Poly(ADP-ribose) polymerase 1(PARP), and promyelocytic leukemia protein(PML) were identified by MALDI-TOF MS. Conclusion: Several proteins were isolated and identified from the 37 mbr-protein complex. Results of this study establish a foundation for further study of the mechanisms by which mbr executes its regulatory function.

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Nan Yang, Yujie Sun, Changyan Ma.[J].南京医科大学学报(自然科学版),2009,29(4):236-240

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  • 收稿日期:2009-04-12
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