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Title: Compositions and methods for making selenocysteine containing polypeptides

Non-naturally occurring tRNA.sup.Sec and methods of using them for recombinant expression of proteins engineered to include one or more selenocysteine residues are disclosed. The non-naturally occurring tRNA.sup.Sec can be used for recombinant manufacture of selenocysteine containing polypeptides encoded by mRNA without the requirement of an SECIS element. In some embodiments, selenocysteine containing polypeptides are manufactured by co-expressing a non-naturally occurring tRNA.sup.Sec a recombinant expression system, such as E. coli, with SerRS, EF-Tu, SelA, or PSTK and SepSecS, and an mRNA with at least one codon that recognizes the anticodon of the non-naturally occurring tRNA.sup.Sec.
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Issue Date:
OSTI Identifier:
Yale University (New Haven, CT) CHO
Patent Number(s):
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Resource Relation:
Patent File Date: 2012 Jul 11
Research Org:
Yale Univ., New Haven, CT (United States)
Sponsoring Org:
Country of Publication:
United States

Works referenced in this record:

An engineered selenocysteine defines a unique class of antibody derivatives
journal, August 2008
  • Hofer, T.; Thomas, J. D.; Burke, T. R.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 34, p. 12451-12456
  • DOI: 10.1073/pnas.0800800105

Sulfur and Selenium: The Role of Oxidation State in Protein Structure and Function
journal, October 2003
  • Jacob, Claus; Giles, Gregory I.; Giles, Niroshini M.
  • Angewandte Chemie International Edition, Vol. 42, Issue 39, p. 4742-4758
  • DOI: 10.1002/anie.200300573

Selenocysteine in proteins—properties and biotechnological use
journal, October 2005
  • Johansson, Linda; Gafvelin, Guro; Arnér, Elias S.J.
  • Biochimica et Biophysica Acta (BBA) - General Subjects, Vol. 1726, Issue 1, p. 1-13
  • DOI: 10.1016/j.bbagen.2005.05.010

Diselenium, instead of disulfide, bonded analogs of conotoxins: novel synthesis and pharmacotherapeutic potential
journal, October 2010

Identification and characterization of a selenoprotein family containing a diselenide bond in a redox motif
journal, August 2007
  • Shchedrina, V. A.; Novoselov, S. V.; Malinouski, M. Yu.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 35, p. 13919-13924
  • DOI: 10.1073/pnas.0703448104

The antioxidant role of selenium and seleno-compounds
journal, May 2003

The many levels of control on bacterial selenoprotein synthesis
journal, November 2009
  • Yoshizawa, Satoko; Böck, August
  • Biochimica et Biophysica Acta (BBA) - General Subjects, Vol. 1790, Issue 11, p. 1404-1414
  • DOI: 10.1016/j.bbagen.2009.03.010

Distinct genetic code expansion strategies for selenocysteine and pyrrolysine are reflected in different aminoacyl-tRNA formation systems
journal, November 2009

RNA-dependent conversion of phosphoserine forms selenocysteine in eukaryotes and archaea
journal, December 2006
  • Yuan, J.; Palioura, S.; Salazar, J. C.
  • Proceedings of the National Academy of Sciences, Vol. 103, Issue 50, p. 18923-18927
  • DOI: 10.1073/pnas.0609703104

Basic local alignment search tool
journal, October 1990
  • Altschul, Stephen F.; Gish, Warren; Miller, Webb
  • Journal of Molecular Biology, Vol. 215, Issue 3, p. 403-410
  • DOI: 10.1016/S0022-2836(05)80360-2

Structurally Modified Firefly Luciferase. Effects of Amino Acid Substitution at Position 286
journal, November 1997
  • Arslan, Tuncer; Mamaev, Sergey V.; Mamaeva, Natalia V.
  • Journal of the American Chemical Society, Vol. 119, Issue 45, p. 10877-10887
  • DOI: 10.1021/ja971927a

Mutagenesis-based definitions and probes of residue burial in proteins
journal, October 2005
  • Bajaj, K.; Chakrabarti, P.; Varadarajan, R.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 45, p. 16221-16226
  • DOI: 10.1073/pnas.0505089102

Positive-selection vectors using the F plasmid ccdB killer gene
journal, October 1994

A method for increasing the yield of properly folded recombinant fusion proteins: Single-chain immunotoxins from renaturation of bacterial inclusion bodies
journal, September 1992
  • Buchner, Johannes; Pastan, Ira; Brinkmann, Ulrich
  • Analytical Biochemistry, Vol. 205, Issue 2, p. 263-270
  • DOI: 10.1016/0003-2697(92)90433-8

A dual-specificity aminoacyl-tRNA synthetase in the deep-rooted eukaryote Giardia lamblia
journal, November 2000
  • Bunjun, S.; Stathopoulos, C.; Graham, D.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 24, p. 12997-13002
  • DOI: 10.1073/pnas.230444397

One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products
journal, May 2000
  • Datsenko, K. A.; Wanner, B. L.
  • Proceedings of the National Academy of Sciences, Vol. 97, Issue 12, p. 6640-6645
  • DOI: 10.1073/pnas.120163297

Global analysis of a “simple” proteome: Methanococcus jannaschii
journal, December 2002
  • Giometti, Carol S.; Reich, Claudia; Tollaksen, Sandra
  • Journal of Chromatography B, Vol. 782, Issue 1-2, p. 227-243
  • DOI: 10.1016/S1570-0232(02)00568-8

Emergence of the universal genetic code imprinted in an RNA record
journal, November 2006
  • Hohn, M. J.; Park, H.-S.; O'Donoghue, P.
  • Proceedings of the National Academy of Sciences, Vol. 103, Issue 48, p. 18095-18100
  • DOI: 10.1073/pnas.0608762103

Toward understanding phosphoseryl-tRNACys formation: The crystal structure of Methanococcus maripaludis phosphoseryl-tRNA synthetase
journal, February 2007
  • Kamtekar, S.; Hohn, M. J.; Park, H.-S.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 8, p. 2620-2625
  • DOI: 10.1073/pnas.0611504104

Cysteinyl-tRNA formation: the last puzzle of aminoacyl-tRNA synthesis
journal, November 1999

Pathogenic mechanism of a human mitochondrial tRNAPhe mutation associated with myoclonic epilepsy with ragged red fibers syndrome
journal, September 2007
  • Ling, J.; Roy, H.; Qin, D.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 39, p. 15299-15304
  • DOI: 10.1073/pnas.0704441104

Possible incorporation of phosphoserine into globin readthrough protein via bovine opal suppressor phosphoseryl-tRNA
journal, October 1986

An aminoacyl-tRNA synthetase that specifically activates pyrrolysine
journal, August 2004
  • Polycarpo, C.; Ambrogelly, A.; Berube, A.
  • Proceedings of the National Academy of Sciences, Vol. 101, Issue 34, p. 12450-12454
  • DOI: 10.1073/pnas.0405362101

Understanding the Sequence Specificity of tRNA Binding to Elongation Factor Tu using tRNA Mutagenesis
journal, March 2009
  • Schrader, Jared M.; Chapman, Stephen J.; Uhlenbeck, Olke C.
  • Journal of Molecular Biology, Vol. 386, Issue 5, p. 1255-1264
  • DOI: 10.1016/j.jmb.2009.01.021

Cysteinyl-tRNA synthetase is not essential for viability of the archaeon Methanococcus maripaludis
journal, November 2001
  • Stathopoulos, C.; Kim, W.; Li, T.
  • Proceedings of the National Academy of Sciences, Vol. 98, Issue 25, p. 14292-14297
  • DOI: 10.1073/pnas.201540498

Protein production by auto-induction in high-density shaking cultures
journal, May 2005

Expanding the Genetic Code of Escherichia coli
journal, April 2001

Molecular genetic studies of a 10.9-kb operon in Escherichia coli for phosphonate uptake and biodegradation
journal, December 1992