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0024968835
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Nonsense suppression is a naturally occurring phenomenon and has been widely used to site-specifically incorporate an unnatural amino acid or analogue into proteins by using an anticodon-adjusted and chemically misacylated suppressor tRNA. Examples are: a) C. J. Noren, S. J. Anthony-Cahill, M. C. Griffith, P. G. Schultz, Science 1989, 244, 182-188;
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0021771662
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c) T. G. Heckler, L. H. Chang, Y. Zama, T. Naka, M. S. Chorghade, S. M. Hecht, Biochemistry 1984, 23, 1468-1473.
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10
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32344450002
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note
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Determined by the band length of the DHFR ORF-UTR domain amplified by PCR from the total cDNA of the E. coli MG1655 strain by using a forward primer specific to the 5′-terminal of the DHFR ORF. Total cDNA was prepared from total mRNA by using a BD SMART mRNA Amplification Kit (BD Biosciences, USA) in combination with Poly(A) Tailing Kit (Ambion, USA).
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11
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0034904102
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Y. Shimizu, A. Inoue, Y. Tomari, T. Suzuki, T. Yokogawa, K. Nishikawa, T. Ueda, Nat. Biotechnol. 2001, 19, 751-755.
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Yokogawa, T.5
Nishikawa, K.6
Ueda, T.7
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12
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32344442610
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note
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There is a small but notable difference in the band intensities for the RT-PCR amplified T7(+)(78) and T7(-)(78) templates.
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13
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4644254373
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The corresponding yields are ≤0.2% (in the presence of anti-ssrA oligonucleotide and protein disulfide isomerase) in the conventional ribosome display by using stop-codon-deleted mRNAs that have a sufficiently long (> 100 aa) 3′ or C-terminal arm. The remarkably enhanced yields obtained here are most likely due to the use of reconstituted translation systems, which are much less contaminated with RNases and other unfavorable components, such as ssrA RNA, and thus protect the mRNA templates from degradation. See, for example: a) A. C. Forster, V. W. Cornish, S. C. Blacklow, Anal. Biochem. 2004, 333, 358-364;
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Forster, A.C.1
Cornish, V.W.2
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0037907668
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b) B.-W. Ying, T. Suzuki, Y. Shimizu, T. Ueda, J. Biochem. 2003, 133, 485-491.
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15
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0030025303
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E. V. Makeyev, V. A. Kolb, A. S. Spirin, FEBS Lett. 1996, 378, 166-170.
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Makeyev, E.V.1
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Spirin, A.S.3
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16
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0034637161
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a) P. Nissen, J. Hansen, N. Ban, P. B. Moore, T. A. Steitz, Science 2000, 289, 920-930;
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Nissen, P.1
Hansen, J.2
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17
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0035805213
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b) M. M. Yusupov, G. Z. Yusupova, A. Baucom, K. Lieberman, T. N. Earnest, J. H. Cate, H. F. Noller, Science 2001, 292, 883-896;
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20
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3643114575
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b) R. Giege, M. Sissler, C. Florentz, Nucleic Acids Res. 1998, 26, 5017-5035.
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Giege, R.1
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21
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0034685609
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a) G. Tocchini-Valentini, M. E. Saks, J. Abelson, J. Mol. Biol. 2000, 298, 779-793;
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Tocchini-Valentini, G.1
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23
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32344452153
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note
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Ser, see ref. [13b].
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24
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32344448224
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note
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Only the T7(+) template was recovered from a 1:1 mixture of T7(+) and T7(-) templates under competitive conditions (Figure S1 in the Supporting Information).
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25
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32344453224
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note
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Ser in read-through of each (ochre, opal, or amber) template (Figure 3c and d) leaves little doubt that Ser is introduced at each stop codon.
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