{"id":574,"date":"2011-04-26T15:21:43","date_gmt":"2011-04-26T19:21:43","guid":{"rendered":"https:\/\/www.bu.edu\/nf-kb\/?page_id=574"},"modified":"2011-04-30T13:57:21","modified_gmt":"2011-04-30T17:57:21","slug":"data-link-11","status":"publish","type":"page","link":"https:\/\/www.bu.edu\/nf-kb\/the-gilmore-lab\/data-link-11\/","title":{"rendered":"Data Link 11"},"content":{"rendered":"

Appendix of Plasmids and Primers for <\/strong><\/h2>\n

Leeman JR, MA Weniger, TF Barth and TD Gilmore. 2008. Deletion analysis and alternative splicing define a transactivation inhibitory domain in human oncoprotein REL. Oncogene 27: 6770-6781<\/p>\n

pGEM-based plasmids<\/h3>\n

pGEM4:<\/span> Cloning vector for in vitro transcription\/translation with either SP6 or T7 promoter elements (Promega)<\/p>\n

pGEM-Hu-cRel:<\/span> XbaI-XhoI\/Klenow fragment of REL subcloned into XbaI-HincII- digested pGEM4 (Barkett et al, 2001)<\/p>\n

pGEM-REL aa 1-322:<\/span> pGEM-Hu-cRel used as a template for PCR amplification to create deletions in RID; A PCR fragment was generated with a 5\u2019 primer located in the SP6 promoter within pGEM-4 and the REL-DRID-R primer located at REL codon 322 containing a novel SalI digestion site. The PCR fragment was digested with XbaI\/SalI and subcloned into pGEM-4 digested with XbaI\/SalI<\/p>\n

pGEM-RELDRID:<\/span> pGEM-Hu-cRel used as a template for PCR amplification; a PCR fragment was generated with 5\u2019 REL-DRID-F forward primer overlapping REL codon 322, also containing a novel SalI site, and a 3\u2019 primer within the T7 promoter of pGEM-4. To generate pGEM RELDRID, the 3\u2019 REL fragment was digested with SalI\/HindIII and subcloned into SalI\/HindIII-digested pGEM-REL aa 1-322<\/p>\n

pGEM-REL-RIDD1: <\/span> pGEM-Hu-cRel was used as a template for PCR amplification; a PCR fragment was generated with a 5\u2019 primer overlapping REL amino acid 356 and containing a novel 5\u2019 SalI site (REL-RIDD1) and a 3\u2019 T7 primer. The fragment was digested with SalI\/HindIII and subcloned into pGEM-RELDRID digested SalI\/HindIII<\/p>\n

pGEM-REL-RIDD1+2:<\/span> pGEM-Hu-cRel used as a template for PCR amplification; a PCR fragment was generated with a 5\u2019 primer overlapping REL codon 389 and containing a novel 5\u2019 SalI site (REL-RIDD2F) and a 3\u2019 T7 primer. The fragment was digested with SalI\/HindIII and subcloned into pGEM-RELDRID digested SalI\/HindIII<\/p>\n

pGEM-REL-RIDD2+3:<\/span> pGEM-Hu-cRel used as a template for PCR amplification; a PCR fragment was generated with a 5\u2019 SP6 primer and a 3\u2019 primer overlapping REL codon 355 and containing a novel SalI site (REL-RIDD2R). The fragment was digested with XbaI\/SalI and subcloned into pGEM-RELDRID digested XbaI\/SalI<\/p>\n

pGEM-REL-RIDD3:<\/span> pGEM-Hu-cRel used as a template for PCR amplification; a PCR fragment was generated with a 5\u2019 SP6 primer and a 3\u2019 primer overlapping REL codon 388 and containing a novel SalI site (REL-RIDD3). The fragment was digested with XbaI\/SalI and subcloned into pGEM-RELDRID digested XbaI\/SalI<\/p>\n

pGEM-RELD164\/VP16:<\/span> REL aa 424-587 replaced with VP16 transactivation sequences. VP16 activation sequences from pSL118 digested with EcoRV\/XhoI and subcloned into Swa\/XhoI-digested pGEM-Hu-c-Rel (D Starczynowski & TD Gilmore, unpublished)<\/p>\n

pGEM-RELD164\/VP16DN:<\/span> VP16 aa 413-453 were removed; pGEM-RELD164\/VP16 was digested with 1) BamHI\/XbaI (to obtain vector sequences); 2) BamHI\/SwaI (to obtain aa 1-423 REL sequences); 3) SmaI\/XbaI (to obtain C-terminal VP16 transactivation domain sequences aa 454-490), resulting in a truncated VP16 transactivation domain fused to RELD164. The three fragments were ligated (D Starczynowski & TD Gilmore, unpublished)<\/p>\n

pGEM-REL<\/span>D<\/span>RID-LacZ-12-106:<\/span> LacZ-12-106 fragment was subcloned as a SalI fragment from pBluescript-SK(+)-LacZ-12-106 into pGEMRELD<\/span>RID digested with SalI.\u00a0 This ligation inserts 95 LacZ codons into REL where RID was removed, along with 4 codons contributed by the flanking SalI sites<\/p>\n

pCRII-based Plasmids<\/strong> <\/span><\/p>\n

pCRII-RELD9:<\/span> cDNA isolated from the Karpas1106 mediastinal B-lymphoma cell line was used as a template for REL<\/em> PCR amplification. A PCR fragment was generated using the 5\u2019 REL-cDNA-F forward primer and the 3\u2019 REL-cDNA-R reverse primer. PCR products were A-tailed and ligated into pCRII<\/p>\n

pCRII-REL+Alu:<\/span> cDNA isolated from the Karpas1106 mediastinal B-lymphoma cell line was used as a template for REL<\/em> PCR amplification. A PCR fragment was generated using the 5\u2019 REL-cDNA-F forward primer and the 3\u2019 REL-cDNA-R reverse primer. PCR products were A-tailed and ligated into pCRII<\/p>\n

pBluescript Intermediate Plasmids <\/strong> <\/span><\/p>\n

pBluescript SK(+):<\/span> Cloning Vector (Stratagene)<\/p>\n

pBluscript SK(+)-REL:<\/span> a wild-type REL EcoRV\/XhoI fragment was subcloned into pBluescript SK(+) digested EcoRV\/XhoI (D Starczynowski & TD Gilmore)<\/p>\n

pBluscript SK(+)-RELD150:<\/span> RELD150 EcoRV\/XhoI fragment subcloned into EcoRV\/XhoI digested pBluescript SK(+) (D Starczynowski & TD Gilmore)<\/p>\n

pBluescript SK(+)-RELD424-490:<\/span> RELD424-490 fragment digested EcoRV\/XhoI was subcloned into pBluescript SK(+) digested with EcoRV\/XhoI (D Starczynowski & TD Gilmore)<\/p>\n

pBluescript SK(+)-RELDRID:<\/span> RELDRID subcloned as an EcoRV\/HindIII fragment into pBluescript SK(+) digested with EcoRV\/HindIII<\/p>\n

pBluescript SK(+)-REL-RIDD1: <\/span> RIDD1 was\u00a0 subcloned as an EcoRV\/HindIII fragment into pBluescript SK(+) digested with EcoRV\/HindIII<\/p>\n

pBluescript SK(+)-REL-RIDD1+2:<\/span> RIDD1+2 subcloned as an EcoRV\/HindIII fragment into pBluescript SK(+) digested with EcoRV\/HindIII<\/p>\n

pBluescript SK(+)-REL-RIDD2+3:<\/span> RIDD2+3 subcloned as an EcoRV\/HindIII fragment into pBluescript SK(+) digested with EcoRV\/HindIII<\/p>\n

pBluescript SK(+)-REL-RIDD3:<\/span> RIDD3 subcloned as an EcoRV\/HindIII fragment into pBluescript SK(+) digested with EcoRV\/HindIII<\/p>\n

pBluescript SK(+)-RELD164\/VP16DN: <\/span> REL aa 427-587 replaced with VP16 activation sequences; pGEM-RELD164\/VP16 digested with EcoRV\/XhoI and subcloned into EcoRV\/XhoI sites in pBluescript SK(+) (D Starczynowski & TD Gilmore) <\/span><\/p>\n

pBluescript SK(+)<\/span>-LacZ-12-106:<\/span> pRSV-bGAL was used as a template for PCR amplification; the 5\u2019 LacZ-R1\/Sal1-F forward primer contained both PCR-generated EcoRI and SalI sites and overlapped LacZ codon 12. The 3\u2019 LacZ-SalI-R reverse primer contained a PCR-generated SalI site and overlapped LacZ codon 106. The final PCR fragment was subcloned as an EcoRI-partial SalI fragment into pBS-SK+<\/p>\n

pSL1180:<\/span> Cloning vector<\/p>\n

pSL1180-VP16:<\/span> VP16 subcloned from CRF3 KS+ (EcoRV\/XbaI) into EcoRV\/XbaI sites of pSL1180 (D Starczynowski & TD Gilmore)<\/p>\n

Avian Retroviral Vectors <\/strong> <\/span><\/h4>\n

JD214BS+:<\/span> Spleen necrosis virus vector (Sif et al, 1993)<\/p>\n

JD-REL:<\/span> JD214BS+ containing human REL subcloned as an XbaI-XhoI fragment into JD214BS+ digested with XbaI-SalI (Gilmore et al, 2001)<\/p>\n

JD-RELDRID:<\/span> pGEM-RELDRID was digested with XbaI\/HindIII and subcloned into JD-REL plasmid digested with XbaI\/HindIII<\/p>\n

JD-RELD424-490:<\/span> JD214BS+ containing human RELD424-490 subcloned as an XbaI\/XhoI fragment into JD214BS+ digested with XbaI\/SalI (Starczynowski et al, 2003)<\/p>\n

JD-RELD9:<\/span> RELD9 XhoI\/HindIII fragment subcloned into JD214 BS+ digested with SalI\/HindIII<\/p>\n

JD-REL+Alu: <\/span> REL+Alu XhoI\/HindIII fragment subcloned into JD214BS+ digested with SalI\/HindIII<\/p>\n

SW253:<\/span> Replication-competent Rev-A avian helper virus (Watanabe & Temin, 1983)<\/p>\n

pcDNA-based Expression Vectors<\/span> <\/strong><\/p>\n

pcDNA 3.1(-):<\/span> CMV promoter-driven expression vector (Invitrogen)<\/p>\n

pcDNA-REL:<\/span> Full-length wild=type REL XbaI\/HindIII fragment subcloned into pcDNA3.1(-) digested with XbaI\/HindIII<\/p>\n

pcDNA-RELDRID:<\/span> RELDRID XbaI\/HindIII fragment subcloned into pcDNA-REL plasmid digested with XbaI\/HindIII<\/p>\n

pcDNA-RELD424-490:<\/span> RELD424-490. RELD424-490 XbaI\/HindIII fragment subcloned into pcDNA3.1(-) digested with XbaI\/HindIII (D Starczynowski & TD Gilmore)<\/p>\n

pcDNA-RELD9:<\/span> A RELD9 XhoI\/BamHI cDNA fragment subcloned into pcDNA3.1(-) digested with XhoI\/BamHI<\/p>\n

pcDNA-REL+Alu:<\/span> A REL+Alu XhoI\/BamHI cDNA fragment subcloned into pcDNA3.1(-) digested with XhoI\/BamHI<\/p>\n

pcDNA-FLAG:<\/span> From Bakary Sylla (World Health Organization, Lyon, France)<\/p>\n

pcDNA-FLAG-RHD-RID:<\/span> pGEM-Hu-cRel was used as a template for the following PCR steps. A PCR fragment was generated using a 5\u2019 primer overlapping REL aa 1 with a novel 5\u2019 EcoRI site and a 3\u2019 primer overlapping REL codon 423 with a novel 3\u2019 BamHI site. The fragment was cloned into pcDNA-FLAG digested with EcoRI\/BamHI (Michael Garbati, Gilmore Lab)<\/p>\n

pcDNA-FLAG-REL:<\/span> pGEM-Hu-cRel was digested EcoRV\/XhoI generating a C-terminal REL fragment that was subcloned into EcoRV\/XhoI-digested pcDNA-FLAG-RHD-RID (Michael Garbati, Gilmore Lab)<\/p>\n

pcDNA-FLAG-RELDRID: <\/span> pcDNA-RELDRID was digested EcoRV\/XhoI generating a C-terminal REL fragment lacking codons 323-422 that was subcloned into EcoRV\/XhoI-digested pcDNA-FLAG-REL<\/p>\n

Expression Vectors for GAL4 Fusion Proteins in A293 Cells <\/span><\/strong><\/h3>\n

<\/strong><\/h2>\n

pSG424<\/span>: Expression vector with the SV40 early promoter controlling the GAL4 DNA-binding domain (aa 1-147) upstream of multi-cloning site (Sadowski & Ptashne, 1989) <\/span><\/p>\n

pSG-REL:<\/span> Wild-type human c-Rel (aa 278-587) fused to GAL4-DBD; pBluescript SK+-REL cut with BamHI-KpnI was subcloned into BamHI\/KpnI-digested pSG424 (Starczynowski et al, 2003)<\/p>\n

pSG-RELD424-490:<\/span> REL (aa 278-423, 491-587) fused to GAL4 DBD; pBluescript SK+RELD424-490 cut with BamHI-KpnI was subcloned into BamHI-KpnI-digested pSG424 (Starczynowski et al., 2003)<\/p>\n

pSG-RELDRID:<\/span> pBluescript SK(+)-RELDRID digested with BamHI-KpnI was subcloned into BamHI-KpnI-digested pSG-REL<\/p>\n

pSG-REL-RIDD1:<\/span> pBluescript SK(+)-REL-RIDD1 digested with BamHI-KpnI was subcloned into BamHI\/KpnI digested pSG424-REL<\/p>\n

pSG-REL-RIDD1+2:<\/span> pBluescript SK(+)-REL RIDD1+2 digested with BamHI-KpnI was subcloned into BamHI\/KpnI-digested pSG-REL<\/p>\n

pSG-REL-RIDD2+3:<\/span> REL RIDD2+3 BamHI-KpnI fragment subcloned into BamHI\/KpnI-digested pSG-REL<\/p>\n

pSG-REL-RIDD3: <\/span> REL RIDD3 BamHI-KpnI fragment subcloned into BamHI\/KpnI- digested pSG-REL<\/p>\n

pSG-RELD150:<\/span> RELD150 (aa 278-436) BamHI-KpnI fragment subcloned into BamHI\/KpnI-digested pSG424 (Starczynowski et al, 2003)<\/p>\n

pSG-VP16DN:<\/span> VP16 transactivation domain sequences were subcloned as an XmaI\/XbaI fragment into the XmaI\/XbaI sites in pSG424<\/p>\n

pSG-REL-RID-VP16DN:<\/span> REL amino acids 424-587 replaced with VP16 activation sequences; pBluescript SK+- REL-RID-VP16DN cut with BamHI-KpnI was subcloned into BamHI\/KpnI-digested pSG424 (also referred to as pSG-3\u2019 RELD164\/VP16; Starczynowski et al, 2003)<\/p>\n

pSG-RELDRID+LacZ: <\/span> pSG–<\/span>RELDRID+LacZ was generated by digestion of pGEM-RELDRID+LacZ with EcoRV\/NdeI and subcloned into pSG-RELD424-490 that had been digested with EcoRV\/NdeI<\/p>\n

pSG-RELD9:<\/span> JD-RELD9 digested with EcoRV\/NdeI was subcloned into EcoRV\/NdeI-digested pSG-REL<\/p>\n

pSG-REL+Alu:<\/span> JD-REL+Alu digested with EcoRV\/NdeI was subcloned into EcoRV\/NdeI-digested pSG-REL<\/p>\n

Yeast GAL4 expression vectors <\/span><\/h3>\n

pGBT9:\u00a0 yeast expression vector for the GAL4 DNA-binding domain (aa 1-147) (Epinat et al, 2000)<\/p>\n

pGB-REL:<\/span> REL (aa 278-587) fused to GAL4-DBD for expression in yeast; pSG424-REL cut with BamHI\/XhoI was subcloned into BamHI\/SalI-digested pGBT9<\/p>\n

pGB-RELDRID:<\/span> RELDRID (from aa 278) BamHI\/XhoI fragment subcloned into BamHI\/SalI-digested pGBT9<\/p>\n

Vertebrate Reporter Plasmids <\/span><\/h3>\n

CMV-bgal:<\/span> CMV promoter-driven expression plasmid for b-galactosidase (Starczynowski et al, 2003, 2005)<\/p>\n

pRSV-bgal:<\/span> Contains the RSV LTR upstream of the b-galactosidase gene (gift of U. Hansen, Boston University)<\/p>\n

pGL2-3x-kB-LMP1-luciferase:<\/span> 3x-kB-luciferase-pGL2 reporter plasmid has a minimal c-fos promoter element and three copies of the major histocompatibility complex (MHC) class I kB element (TGGGGATTCCCCA) upstream of\u00a0 the luciferase gene (Mitchell & Sugden, 1995; gift of G. Mosialos, Fleming Institute)<\/p>\n

IkB-pGL2 luciferase:<\/span> 1.3 kb HindIII-EcoRI fragment of chicken IkBa, containing the transcriptional start site and 900 bp of upstream sequence all upstream of the luciferase gene (Schatzle et al, 1995; Starczynowski et al, 2007)<\/p>\n

PolyA-GAL4E1b GAL4-luc:<\/span> Contains GAL4 DNA-binding sites upstream of the luciferase gene (Starczynowski et al, 2003). Gift of Joseph Lipsick (Stanford Medical School)<\/p>\n

Primers used for PCR<\/span><\/strong> <\/span><\/strong><\/h3>\n

SP6 Promoter:<\/span> 5\u2019-GATTTAGGTGACACTATAG-3\u2019<\/p>\n

T7 Promoter: <\/span> 5\u2019-GTAATACGACTCACTATAGGGC-3\u2019<\/p>\n

REL-423-R:<\/span> 5\u2019-CGCAGGATCC<\/span>CAATCATTCCCAACAGG-3\u2019<\/p>\n

PCR generated BamHI site (GGATCC): underlined<\/p>\n

RELDRID-F:<\/span> 5\u2019-GGGGTCGAC<\/span>TTAAATGCTTCTAATGC-3\u2019<\/p>\n

RELDRID-R:<\/span> 5\u2019-TCCGTCGAC<\/span>AATTGAACCGAGGA-3\u2019<\/p>\n

REL-RIDD1:<\/span> 5\u2019-AATCGTCGAC<\/span>TATCCCTCACCTGGGCCCATC-3\u2019<\/p>\n

REL-RIDD3:<\/span> 5\u2019-AATAGTCGAC<\/span>TGGGGTGGGGTGGGCCAC-3\u2019<\/p>\n

REL-RIDD2R:<\/span> 5\u2019-ATACGTCGAC<\/span>GTAGGATTCTGCTTGACTTGAAACC-3\u2019<\/p>\n

REL-RIDD2F:<\/span> 5\u2019-ACGCGTCGAC<\/span>CGCTCAGGCAATACAAACCCACTG-3\u2019<\/p>\n

PCR generated SalI sites (GTCGAC): underlined<\/p>\n

LacZ-R1\/Sal1-F:<\/span> 5\u2019-AAGAATTC<\/span>GTCGAC<\/span>TTACAACGTCGTGACTGGGAAAAC-3\u2019<\/p>\n

PCR generated EcoRI site (GAATTC) and SalI site (GTCGAC): underlined<\/p>\n

LacZ-Sal1-R:<\/span> 5\u2019-TAATGTCGAC<\/span>ATAGGTTACGTTGGTGTAGATGGG-3\u2019<\/p>\n

PCR generated SalI site (GTCGAC): underlined<\/p>\n

REL-cDNA-F:<\/span> 5′-CACCATGGCCTCCGGTGCGTATA-3′<\/p>\n

REL-cDNA-R:<\/span> 5′-TTATACTTGAAAAAATTCATATGGAAAGGAGTC-3′<\/p>\n

Primers used for Real-time PCR <\/span><\/h3>\n

GAPDH-For:<\/span> 5\u2019-TGGTATCGTGGAAGGACTCATGAC-3\u2019 <\/span><\/p>\n

GAPDH-Rev:<\/span> 5\u2019-ATGCCAGTGAGCTTCCCGTTCAGC-3\u2019 <\/span><\/p>\n

REL-558-Rev:<\/span> 5\u2019-CCATGACTGTTTGGATTAGTACTGTTTG-3\u2019 <\/span><\/p>\n

REL-AltSplic-d9-For:<\/span> 5\u2019-AAACTGTGCCAGGATCACGAACC-3\u2019 <\/span><\/p>\n

REL-AltSplice-For:<\/span> 5\u2019-GCTATCACAGAACCCGTAACAG-3\u2019 <\/span><\/p>\n

REL-AltSplice-Rev:<\/span> 5\u2019-ACCCCTGTAGGCATTTCTCTCACA-3\u2019 <\/span><\/p>\n

Oligonucleotides used for kB probe <\/strong> <\/span><\/p>\n

IFN-b kB-site:<\/span> 5\u2019-TCGAGAGGTCGGGAAATTCC<\/span>CCCCCG-3\u2019<\/p>\n

MHC kB-site:<\/span> 5\u2019-TCGAGAGGTTGGGGATTCCCCA<\/span>CCCG-3\u2019<\/p>\n

SOD2 kB-site:\u00a0 5\u2019-TCGAGAGGTCGGGAATACCC<\/span>CCCCCG-3\u2019 (Bernard et al, 2001)<\/p>\n

kB site underlined<\/p>\n

<\/h3>\n

References<\/strong><\/h3>\n

Abid MR, IG Schoots, KC Spokes, SQ Wu, C Mawhinney & WC Aird (2004) Vascular endothelial growth factor-mediated induction of manganese superoxide dismutase occurs through redox-dependent regulation of forkhead and IkB\/NF-kB. Journal of Biological Chemistry 279: 44030-44038<\/p>\n

Barkett M, JE Dooher, L Lemonnier, L Simmons, JN Scarpati, Y Wang & TD Gilmore (2001) Three mutations in the retroviral oncoprotein v-Rel render it resistant to cleavage by caspase-3. Biochimica et Biophysica Acta 1526: 25-36<\/p>\n

Bernard D, B Quatannens, A Begue, B Vandenbunder & C Abbadie (2001) Antiproliferative and antiapoptotic effects of cRel may occur witin the same cells via the up-regulation of manganese superoxide dismutase. Cancer Research 61: 2656-2664<\/p>\n

Epinat J-C, D Kazandjian, DD Harkness, S Petros, J Dave, DW White & TD Gilmore (2000) Mutant envelope residues confer a transactivation function onto N-terminal sequences of the v-Rel oncoprotein. Oncogene 19:<\/strong> 599-607<\/p>\n

Gilmore TD, C Cormier, J Jean-Jacques & M-E Gapuzan (2001) Malignant transformation of primary chicken spleen cells by human transcription factor c-Rel. Oncogene 20: 7098-7103<\/p>\n

Kalaitzidis D, RE Davis, A Rosenwald, LM Staudt & TD Gilmore (2002) The human B-cell lymphoma cell line RC-K8 has multiple genetic alterations that dysregulate the Rel\/NF-kB signal transduction pathway. Oncogene 21: 8759-8768<\/p>\n

Mitchell T & B Sugden\u00a0 (1995) Stimulation of NF-kB-mediated transcription by mutant derivatives of the latent membrane protein of Epstein-Barr virus. Journal of Virology 69: 2968-2976<\/p>\n

Sadowski I & M Ptashne (1989) A vector for expressing GAL4(1-147) fusions in mammalian cells. Nucleic Acids Research 17: 7539<\/p>\n

Schatzle JD, J Kralova & HR Bose Jr (1995) Avian IkBa is transcriptionally induced by c-Rel and v-Rel with different kinetics. Journal of Virology 69: 5383-5390<\/p>\n

Sif S, AJ Capobianco & TD Gilmore (1993) The v-Rel oncoprotein increases expression from Sp1 site-containing promoters in chicken embryo fibroblasts. Oncogene 8: 2501-2509<\/p>\n

Starczynowski DT, JG Reynolds & TD Gilmore. (2003) Deletion of either C-terminal transactivation subdomain enhances the in vitro transforming activity of human transcription factor REL in chicken spleen cells. Oncogene 22: 6928-6936<\/p>\n

Starczynowski DT, JG Reynolds & TD Gilmore (2005) Mutations of tumor necrosis factor alpha-responsive serine residues within the C-terminal transactivation domain of human transcription factor REL enhance its in vitro transforming ability. Oncogene 24: 7355-7368<\/p>\n

Starczynowski DT, H Trautmann, C Pott, L Harder, N Arnold, JA Africa, JR Leeman, R Siebert & TD Gilmore (2007) Mutation of an IKK phosphorylation site within the transactivation domain of REL in two patients with B-cell lymphoma enhances REL’s in vitro transforming activity. Oncogene 26: 2685-2694<\/p>\n

Watanabe S & HM Temin (1983) Construction of a helper cell line for avian reticuloendotheliosis virus cloning vectors. Molecular and Cellular Biology 3: 2241-2249<\/p>\n","protected":false},"excerpt":{"rendered":"

Appendix of Plasmids and Primers for Leeman JR, MA Weniger, TF Barth and TD Gilmore. 2008. Deletion analysis and alternative splicing define a transactivation inhibitory domain in human oncoprotein REL. Oncogene 27: 6770-6781 pGEM-based plasmids pGEM4: Cloning vector for in vitro transcription\/translation with either SP6 or T7 promoter elements (Promega) pGEM-Hu-cRel: XbaI-XhoI\/Klenow fragment of REL […]<\/p>\n","protected":false},"author":4258,"featured_media":0,"parent":337,"menu_order":13,"comment_status":"closed","ping_status":"open","template":"","meta":[],"_links":{"self":[{"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages\/574"}],"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=574"}],"version-history":[{"count":3,"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages\/574\/revisions"}],"predecessor-version":[{"id":622,"href":"https:\/\/www.bu.edu\/nf-kb\/wp-json\/wp\/v2\/pages\/574\/revisions\/622"}],"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=574"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}