Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (256K) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Right arrow Commercial Re-use Guidelines
for Open Access NAR Content
Google Scholar
Right arrow Articles by Kurz, M.
Right arrow Articles by Lohse, P. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kurz, M.
Right arrow Articles by Lohse, P. A.
Related Collections
Right arrow Protein-nucleic acid interaction
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2000, Vol. 28, No. 18 e83
© 2000 Oxford University Press

Psoralen photo-crosslinked mRNA–puromycin conjugates: a novel template for the rapid and facile preparation of mRNA–protein fusions

Markus Kurz, Ke Gu and Peter A. Lohse*

Phylos Inc., 128 Spring Street, Lexington, MA 02421, USA

We describe the synthesis of a novel type of mRNA template and its use in the preparation of mRNA–protein fusions. A light-induced psoralen crosslinking reaction was used to attach a puromycin-containing oligonucleotide to the 3'-end of an mRNA template. The photo-crosslinked template was found to undergo efficient mRNA–protein fusion formation in rabbit reticulocyte lysate. Fusion formation was subsequently tested with templates carrying puromycin linkers of different length and chemical composition. Short linkers with multiple triethyleneglycol phosphate building blocks allowed the most efficient fusion formation under a wide range of salt conditions. The present method simplifies the preparation of mRNA–protein fusions and thus significantly accelerates the in vitro protein evolution procedure which involves repetitive cycles of fusion production and selection.

* To whom correspondence should be addressed. Tel: +1 781 862 6400; Fax: +1 781 402 8800; Email: plohse@phylos.com


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Nucleic Acids ResHome page
J. Yamaguchi, M. Naimuddin, M. Biyani, T. Sasaki, M. Machida, T. Kubo, T. Funatsu, Y. Husimi, and N. Nemoto
cDNA display: a novel screening method for functional disulfide-rich peptides by solid-phase synthesis and stabilization of mRNA-protein fusions
Nucleic Acids Res., September 1, 2009; 37(16): e108 - e108.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Biyani, Y. Husimi, and N. Nemoto
Solid-phase translation and RNA-protein fusion: a novel approach for folding quality control and direct immobilization of proteins using anchored mRNA
Nucleic Acids Res., November 6, 2006; 34(20): e140 - e140.
[Abstract] [Full Text] [PDF]

Home page
 Protein Eng Des SelHome page
W. W. Ja, B. N. Olsen, and R. W. Roberts
Epitope mapping using mRNA display and a unidirectional nested deletion library
Protein Eng. Des. Sel., July 1, 2005; 18(7): 309 - 319.
[Abstract] [Full Text] [PDF]

Home page
 Proc. Natl. Acad. Sci. USAHome page
X. Shen, C. A. Valencia, J. W. Szostak, B. Dong, and R. Liu
Scanning the human proteome for calmodulin-binding proteins
PNAS, April 26, 2005; 102(17): 5969 - 5974.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
E. Miyamoto-Sato, H. Takashima, S. Fuse, K. Sue, M. Ishizaka, S. Tateyama, K. Horisawa, T. Sawasaki, Y. Endo, and H. Yanagawa
Highly stable and efficient mRNA templates for mRNA-protein fusions and C-terminally labeled proteins
Nucleic Acids Res., August 1, 2003; 31(15): e78 - e78.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.