{"title":"bcl2基因主断点区(mbr)结合蛋白的分离与鉴定","authors":"Nan Yang, Yujie Sun, Changyan Ma","doi":"10.1016/S1007-4376(09)60062-9","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>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.</p></div><div><h3>Methods</h3><p>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.</p></div><div><h3>Results</h3><p>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.</p></div><div><h3>Conclusion</h3><p>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.</p></div>","PeriodicalId":100807,"journal":{"name":"Journal of Nanjing Medical University","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2009-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1007-4376(09)60062-9","citationCount":"0","resultStr":"{\"title\":\"Isolation and identification of proteins binding to the major breakpoint region(mbr) of bcl2 gene\",\"authors\":\"Nan Yang, Yujie Sun, Changyan Ma\",\"doi\":\"10.1016/S1007-4376(09)60062-9\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><p>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.</p></div><div><h3>Methods</h3><p>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.</p></div><div><h3>Results</h3><p>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.</p></div><div><h3>Conclusion</h3><p>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.</p></div>\",\"PeriodicalId\":100807,\"journal\":{\"name\":\"Journal of Nanjing Medical University\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2009-07-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1016/S1007-4376(09)60062-9\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Nanjing Medical University\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1007437609600629\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Nanjing Medical University","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1007437609600629","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
Isolation and identification of proteins binding to the major breakpoint region(mbr) of bcl2 gene
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.