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Nucl. Acids Res. 34, e7. Four-base codon mediated mRNA display to construct peptide libraries that contain multiple nonnatural amino acids. 2006

Muranaka, N., Hohsaka, T. and Sisido, M.

Notes: The authors devised an mRNA display system to generate a peptide library with multiple nonnatural amino acids incorporated into the proteins, an important feature of peptide libraries for successful drug discovery. An mRNA with 3 four-base codons at a random position was used as a template in an in vitro translation system in the presence of charged tRNAs carrying four-base codons. In vitro translations were performed using 3.6 × 1013 molecules of mRNA template and the E. coli S30 Extract System. The mRNA template contained a T7 tag sequence, so the translation products could be detected using an anti-T7 tag antibody and the Anti-Mouse IgG (H+L), AP Conjugate. The mRNA-displayed peptides also incorporated a polyhistidine tag so that they could be purified using the MagneHis™ Ni-Particles. After selecting for the desired protein characteristic, the mRNA portion of the mRNA-displayed peptides was reverse transcribed and quantitated by real-time PCR. PCR products were cloned into the pGEM®-T Vector prior to sequencing. (3651)

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Nat. Neurosci. 3, 323-29. Synapsins as mediators of BDNF-enhanced neurotransmitter release. 2000

Jovanovic, J.N., Czernik, A.J., Fienberg, A.A., Greengard, P., and Sihra, T.S.

Notes: Treatment of  brain-derived neurotrophic factor increased MAPK-dependent synapsin I phosphorylation in rat cerebral cortex synaptosomal preparations. MAPK activity was determined by Western blot analysis using the Anti-ACTIVE® MAPK (1:10,000 dilution) pAb to detect the dually phosphorylated forms of MAPK. CaM kinase II activity was assayed by Western blot analysis with the Anti-ACTIVE® CaM KII pAb, Rabbit, (pT286) (1:2500 dilution). The Anti-Mouse IgG (H+L), AP Conjugate (1:1000 dilution) was used as a secondary antibody in Western blot analysis to detect TrkB expression. (2393)

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