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J. Biol. Chem. 282, 29144-29151. The membrane topology of RTN3 and its effect on binding of RTN3 to BACE1 2007

He, W., Shi, Q., Hu, X. and Yan, R.

Notes: The authors of this study determined the membrane topology of reticulon 3 (RTN3), an integral membrane protein that is expressed at high levels in neruons and has been show to negatively regulate the activity of BACE1 (Beta site APP-Cleaving Enzyme). Disruption of RTN3 is associated with incidence of dystrophic neurites in AD brain. RTN3 was translated using the TNT® Quick Coupled Transcription/Translation System in the presence of Canine Microsomal Membranes and labeled using the Transcend™ Non-Radioactive Translation Detection System.

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J. Virol. 80, 6669-6677. Human papillomavirus e7 oncoprotein dysregulates steroid receptor coactivator 1 localization and function.

Baldwin, A., Huh, K-W. and Mϋnger, K.

Notes: The MagneGST™ Protein Purification System was used to purify GST fusion proteins of the oncoprotein HPV16 E7 or various mutants of HPV16 E7. The purified GST fusion proteins were used for in vitro binding experiments with steroid receptor coactivator 1 (SRC-1), which was produced using the TNT® T7 Coupled Wheat Germ Extract System and labeled with the Transcend™ Non-Radioactive Translation Detection System. GST pull-down assays were resolved by Western analysis using streptavidn-horseradish peroxidase and alpha-GST. To determine the effects of endogenously expressed HPV16 E7 on SCR-1-mediated transcription, luciferase reporters under the control of either the IL-8 promoter or an artificial promoter containing three estrogen response elements repeats (3 × EREs) were cotransfected with a Renilla control vector into two human cervical cancer lines (C33A and CaSki) using either the Transfast™ Transfection Reagent or another commercial transfection reagent. The Dual-Luciferase® Reporter Assay was then used to determine luciferase activity to functionally map the E7-interacting domain and to determine the effects of high- and low-risk PHV E7s on SRC-1-mediated transcription. (3459)

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J. Med. Genet. 38, 102-105. De novo recurrent germline mutation of the BRCA2 gene in a patient with early onset breast cancer. 2001

van der Luijt, R.B., van Zon, P.H., Jansen, R.P., van der Sijs-Bos, C.J., Warlam-Rodenhuis, C.C. and Ausems, M.G.

Notes: This paper describes using the TNT® T7 Coupled Reticulocyte Lysate System and Transcend™ tRNA to perform a nonradioactive in vitro Protein Truncation Test for a BRCA2 gene product.  Templates for the transcription-translation reactions were created by PCR-amplifying regions of the BRCA2 gene from normal and cancer patient sample genomic DNA.  (3075)

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J. Biol. Chem. 275, 1176-1182. Phosphorylation modulates the function of the vasoactive intestinal polypeptide receptor transcriptional repressor protein. 2000

Pei, L.

Notes: The TNT® Coupled Reticulocyte Lysate System was used in conjunction with the Transcend™ Chemiluminescent Non-Radioactive Translation Detection System. Various truncations of the protein of interest were produced ranging from 67kDa to 19kDa to use in gel shift assays. The truncations were used to identify which region bound the oligo of interest. The in vitro translated proteins were also used with antibodies for supershifts. (0003)

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J. Biol. Chem. 274, 3151-3158. Pituitary tumor-transforming gene protein associates with ribosomal protein S10 and a novel human homologue of DnaJ in testicular cells. 1999

Pei, L.

Notes: The authors used the TNT® Coupled Reticulocyte Lysate System with Transcend™ Biotinylated tRNA to synthesize S10 and HSJ2 protein for protein:protein interaction studies with GST-tagged PTTG. (0556)

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J. Biol. Chem. 274, 15671-15677. Splice variants of intersectin are components of the endocytic machinery in neurons and nonneuronal cells. 1999

Hussain, N.K., Yamabhai, M., Ramjaun, A.R., Guy, A.M., Baranes, D., O'Bryan, J.P., Der, C.J., Kay, B.K., McPherson, P.S.

Notes: The proteins Ibp2, MP90 and luciferase were translated in vitro with the TNT® Coupled Reticulocyte Lysate System in the presence of the Transcend™ Biotinylated tRNA. The translated proteins were reacted with a glutathione bead-immobilized portion of the clathrin heavy chain. Bound proteins were solubilized in SDS sample buffer and the proteins detected by Western detection with the Transcend™ Non-Radioactive Translation Detection System. (0990)

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J. Biol. Chem. 271, 14800-14806. The major astrocytic phosphoprotein PEA-15 is encoded by two mRNAs conserved on their full length in mouse and human. 1996

Estelles, A. , Yokoyama, M. , Nothias, F. , Vincent, J. D. , Glowinski, J. , Vernier, P. , Chneiweiss, H.

Notes: PEA-15 cDNAs were cloned from a mouse astrocytic library, and the clones were linearized and expressed using T3 or T7 Polymerase in the TNT® Reticulocyte Lysate System either in the presence of [35S]methionine or Transcend™ tRNA. The resulting proteins were analyzed by SDS-PAGE and Western blotting, respectively. A unique 15kDa protein that was recognized by an antibody to PEA-15 was produced in the TNT® System. (1178)

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