Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow Print PDF (527K) 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 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 He, M.
Right arrow Articles by Taussig, M. J.
Right arrow Search for Related Content
PubMed
Right arrow Articles by He, M.
Right arrow Articles by Taussig, M. J.
Related Collections
Right arrow New restriction enzymes
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Nucleic Acids Research, 2001, Vol. 29, No. 15 e73
© 2001 Oxford University Press

Single step generation of protein arrays from DNA by cell-free expression and in situ immobilisation (PISA method)

Mingyue He* and Michael J. Taussig

Technology Research Group, The Babraham Institute, Babraham, Cambridge CB2 4AT, UK

We describe a format for production of protein arrays termed ‘protein in situ array’ (PISA). A PISA is rapidly generated in one step directly from PCR-generated DNA fragments by cell-free protein expression and in situ immobilisation at a surface. The template for expression is DNA encoding individual proteins or domains, which is produced by PCR using primers designed from information in DNA databases. Coupled transcription and translation is carried out on a surface to which the tagged protein adheres as soon as it is synthesised. Because proteins generated by cell-free synthesis are usually soluble and functional, this method can overcome problems of insolubility or degradation associated with bacterial expression of recombinant proteins. Moreover, the use of PCR-generated DNA enables rapid production of proteins or domains based on genome information alone and will be particularly useful where cloned material is not available. Here we show that human single-chain antibody fragments (three domain, VH/K form) and an enzyme (luciferase) can be functionally arrayed by the PISA method.

* To whom correspondence should be addressed. Tel: +44 1223 496249; Fax: +44 1223 496045; Email: mingyue.he{at}bbsrc.ac.uk


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
 Genome ResHome page
Y. Sasuga, T. Tani, M. Hayashi, H. Yamakawa, O. Ohara, and Y. Harada
Development of a microscopic platform for real-time monitoring of biomolecular interactions
Genome Res., January 1, 2006; 16(1): 132 - 139.
[Abstract] [Full Text] [PDF]

Home page
 J Biomol ScreenHome page
T. Vanhercke, C. Ampe, L. Tirry, and P. Denolf
Rescue and In Situ Selection and Evaluation (RISE): A Method for High-Throughput Panning of Phage Display Libraries
J Biomol Screen, March 1, 2005; 10(2): 108 - 117.
[Abstract] [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.