Notes: A fusion protein consisting of the HA epitope fused to APC (adenomatous polyposis coli) and APC deletion mutant proteins were expressed using the TNT® T7 Coupled in vitro Transcription/Translation System. In coprecipitation experiments, N-terminal fragments were able to form complexes with HA-APC. Also, APC fragments did not associate with HA-APC when they were translated separately and then mixed together. The first 171 residues of the APC protein were sufficient for complex formation. (1849)

Notes: Specificity of the GST-Cdi1 antisera was demonstrated by immunoprecipitation of Cdi1 protein produced in the TNT^® Coupled Reticulocyte Lysate System. (1795)

Notes: Pu.1, the macrophage- and B-cell-specific ets-related transcription factor, was synthesized using the TNT^® T7 Coupled Reticulocyte Lysate System. In electrophoretic mobility shift assays Pu.1 binds to PuB2, one of two purine-rich binding sites in the HIV-2 enhancer. (1803)

Notes: Early activation antigen CD69 was expressed using the TNT^® Coupled Reticulocyte Lysate System. Cotranslational processing was investigated using Canine Pancreatic Microsomal Membranes. The two cotranslationally processed products of 28kDa and 34kDa are the different glycosylated forms of the protein. (1057)

Notes: Both p105 and p98 precursors were co-translated in the TNT^® Coupled Reticulocyte Lysate System and their binding to NF-KB was determined. (0682)

Notes: TATA-binding protein (TBP) and p53 were synthesized using the TNT^® Coupled Reticulocyte Lysate System. TBP coimmunoprecipitates with both wild type and mutant p53. The complex of TBP and p53 also binds the TATA box. The success of transcription may depend more on the conformation of the TBP-p53 complex than just p53 binding to TBP. (0737)

Notes: RNA isolated from peripheral blood lymphocytes of DMD patients was used as a template for RT-PCR of a region of the dystrophin gene. The protein truncation test was performed using proteins transcribed and translated from the RT-PCR products with the TNT^® T7 Coupled Reticulocyte Lysate System. The translation products were analyzed by SDS-PAGE. PTT detects nonsense but not missense mutations. (0492)

Notes: PTT was performed on the RNA of a DMD patient and his mother, a carrier of the DMD mutation. Total RNA was isolated from blood. RT-PCR and 'boosted' PCR was used to amplify a partial cDNA of the gene with a T7 promoter. The engineered template was expressed in vitro with the TNT^® T7 Coupled Reticulocyte Lysate System in the presence of [³H]leucine followed by SDS-PAGE. The PTT may be used to screen the entire coding region of the DMD gene in 5-10 reactions using 5 overlapping primer sets for the primary PCR and 10 nested primer sets for the 'boosted' PCR. (0493)

Notes: The aryl hydrocarbon receptor nuclear translocator protein (ARNT) was generated using the TNT® T7 Coupled Reticulocyte Lysate System. The ARNT protein was added to ARNT-deficient cytosolic extracts. ARNT associated with the Ah receptor and restored Ah receptor-dependent xenobiotic-responsive element (XRE)-binding activity. (1831)

Notes: [³⁵S]methionine-labeled bcl-3 proteins were generated using the TNT^® T3 or T7 Coupled Reticulocyte Lysate System. bcl-3 was immunoprecipitated with anti-bcl-3 but not with anti-pp40 serum. The bcl-3 protein was also subjected to complete digestion with trypsin, oxidation of soluble peptides and lyophilization before high-voltage electrophoresis. The bcl-3 protein is related to the human 37kDa (IkappaBalpha) protein. (1764)