{"id":556,"date":"2011-04-26T15:09:12","date_gmt":"2011-04-26T19:09:12","guid":{"rendered":"https:\/\/www.bu.edu\/nf-kb\/?page_id=556"},"modified":"2011-04-28T19:02:41","modified_gmt":"2011-04-28T23:02:41","slug":"data-link-4","status":"publish","type":"page","link":"https:\/\/www.bu.edu\/nf-kb\/the-gilmore-lab\/data-link-4\/","title":{"rendered":"Data Link 4"},"content":{"rendered":"<p><strong>APPENDIX<\/strong><\/p>\n<p>Presented below is a more detailed version of plasmid constructions for recombinant DNA molecules used in:<\/p>\n<p>Gilmore TD, C Cormier, J Jean-Jacques &amp; M-E Gapuzan. (2001) Malignant transformation of primary chicken spleen cells by human transcription factor c-Rel. Oncogene 20: 7098-7103<\/p>\n<p>Further details can be obtained by sending a request for information to Tom Gilmore at gilmore@bu.edu.<\/p>\n<p><strong>Plasmid descriptions and constructions<\/strong><\/p>\n<p>\u00b7      Plasmid pMH105 (v-Rel\/Neo) was a kind gift of Mark Hannink (University of Missouri) and has been described previously (White &amp; Gilmore, 1996).<\/p>\n<p>\u00b7      Plasmid c-Rel\/Neo (human c-rel and neo) was constructed by subcloning an XbaI to SpeI fragment containing a full-length human c-rel cDNA into pMH105 which had been cleaved with XbaI.This excises v-rel and thus human c-rel replaces v-rel as the 5\u2019 gene.<\/p>\n<p>\u00b7      The plasmid for only the expression of Bcl-2 was created by digesting plasmid v-R273H-bcl-2 (White &amp; Gilmore, 1996) with XbaI and religating (this removes the 5\u2019 mutant v-rel gene).<\/p>\n<p>\u00b7      Plasmid c-Rel\/Bcl-2 was made by inserting the XbaI-SpeI fragment containing human c-rel (see above) at the XbaI site of the Bcl-2 (alone) expression plasmid.<\/p>\n<p>\u00b7      In plasmid c-Rel\/Bcl-XL the human bcl-xL cDNA was used to replace the 3\u2019 neo gene (removed by NcoI\/RsrII digestion from pMH105 [i.e., v-Rel\/Neo]).This created a v-Rel\/Bcl-xL vector.This vector was then digested with XbaI (to remove the 5\u2019 v-rel gene) and v-rel was replaced by an XbaI-SpeI fragment containing human c-rel.<\/p>\n<p>\u00b7      Plasmid c-Rel\/mBcl-2 contains a mutant chicken bcl-2 gene, which is lacking the BH4 domain, and was made by digesting the bcl-2 gene with NaeI and SmaI and religating.These blunt-end enzymes result in the in-frame deletion of codons 4 to 33 of chicken bcl-2.<\/p>\n<p><strong>References<\/strong><\/p>\n<p>DW White &amp; TD Gilmore (1996) Oncogene 13, 891-899.<\/p>\n","protected":false},"excerpt":{"rendered":"<p>APPENDIX Presented below is a more detailed version of plasmid constructions for recombinant DNA molecules used in: Gilmore TD, C Cormier, J Jean-Jacques &amp; M-E Gapuzan. (2001) Malignant transformation of primary chicken spleen cells by human transcription factor c-Rel. Oncogene 20: 7098-7103 Further details can be obtained by sending a request for information to Tom [&hellip;]<\/p>\n","protected":false},"author":4258,"featured_media":0,"parent":337,"menu_order":5,"comment_status":"closed","ping_status":"open","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages\/556"}],"collection":[{"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/users\/4258"}],"replies":[{"embeddable":true,"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/comments?post=556"}],"version-history":[{"count":3,"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages\/556\/revisions"}],"predecessor-version":[{"id":1010,"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages\/556\/revisions\/1010"}],"up":[{"embeddable":true,"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages\/337"}],"wp:attachment":[{"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/media?parent=556"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}