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Proc. Natl. Acad. Sci. USA 95, 15803-15808. Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord. 1998

Coulouarn, Y. , Lihrmann, I. , Jegou, S. , Anouar, Y. , Tostivint, H. , Beauvillain, J. C. , Conlon, J. M. , Bern, H. A. , Vaudry, H.

Notes: In this paper, Tfl DNA polymerase was used for two-step RT-PCR. The amplified fragments were cloned into the pGEM®-T Vector. Promega's Terminal Deoxynucleotidyl Transferase (TdT) was used to end label specific oligos for screening a frog brain cDNA library. T7 and T3 RNA polymerases were used to make digoxigenin labeled riboprobes for in situ hybridization. (1281)

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Biochemistry 32, 83-90. Membrane translocation of diphtheria toxin A-fragment: Role of carboxy-terminal region. 1993

Ariansen, S., Afanasiev, B.N., Moskaug, J.O., Stenmark, H., Madshus, I.H. and Olsnes, S.

Notes: Enzymatically active diphtheria toxin was formed in vitro by transcription/translation from PCR products carrying the A- and B-fragments of the toxin. T3 RNA Polymerase was used for in vitro transcription and the Rabbit Reticulocyte Lysate System was used for translation of proteins. Both wild type and mutant proteins were produced and assayed. The synthesized proteins were tested for trypsin sensitivity, ability to inhibit protein synthesis, binding and translocation in Vero cells. Considerable alterations can be made to the C-terminal region of the protein without blocking translocation. Some mutant toxins were less toxic than wild type toxins in unnicked form, but when nicked most exhibited the same toxicity. (2226)

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