Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency.

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency. Research Abstract Details 

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Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency. Abstract Text:

Erik A Rodriguez,Henry A Lester,Dennis A Dougherty,

The incorporation of unnatural amino acids into proteins is a valuable tool for addition of biophysical probes, bio-orthogonal functionalities, and photoreactive cross-linking agents, although these approaches often require quantities of protein that are difficult to access with chemically aminoacylated tRNAs. THG73 is an amber suppressor tRNA that has been used extensively, incorporating over 100 residues in 20 proteins. In vitro studies have shown that the Escherichia coli Asn amber suppressor (ENAS) suppresses better than THG73. However, we report here that ENAS suppresses with <26% of the efficiency of THG73 in Xenopus oocytes. We then tested the newly developed Tetrahymena thermophila Gln amber suppressor (TQAS) tRNA library, which contains mutations in the second to fourth positions of the acceptor stem. The acceptor stem mutations have no adverse effect on suppression efficiency and, in fact, can increase the suppression efficiency. Combining mutations causes an averaging of suppression efficiency, and increased suppression efficiency does not correlate with increased DeltaG of the acceptor stem. We created a T. thermophila opal suppressor, TQOpS', which shows approximately 50% suppression efficiency relative to THG73. The TQAS tRNA library, composed of functional suppressor tRNAs, has been created and will allow for screening in eukaryotic cells, where rapid analysis of large libraries is not feasible.

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency. Publishing Authors By Initials

EA Rodriguez,HA Lester,DA Dougherty,

For similar animals: invertebrates: protozoa: ciliophora: oligohymenophorea: hymenostomatida: tetrahymenina: tetrahymena: tetrahymena thermophila research abstracts see: animals: invertebrates: protozoa: ciliophora: oligohymenophorea: hymenostomatida: tetrahymenina: tetrahymena: tetrahymena thermophila research

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Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: RNA (New York, N.Y.)

VOLUME: 13

Page Numbers: 1715-22

Journal Abbreviation: RNA

ISSN: 1355-8382

DAY: 13

MONTH: 08

YEAR: 2007

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 9509184

Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency. Keywords Mesh Terms:

KEYWORDS: Tetrahymena thermophila

MESH TERMS: metabolism

Chemical & Substance for Abstract: Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency. Information

Substance Name: RNA, Transfer

Registry Number: 9014-25-9

Grant and Affiliation Information for Improved amber and opal suppressor tRNAs for incorporation of unnatural amino acids in vivo. Part 2: evaluating suppression efficiency.

AFFILIATION: Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.

Country: United States

AGENCY: United States NINDS

GRANT: NS 34407

ACRONYM: NS

MEDLINETA: RNA

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