PCR-linked in vitro expression: a novel system for high-throughput construction and screening of protein libraries

被引：44

作者：

Rungpragayphan, S ^{[1
]}

Nakano, H ^{[1
]}

Yamane, T ^{[1
]}

机构：

[1] Nagoya Univ, Grad Sch Biol & Agr Sci, Lab Mol Biotechnol, Chikusa Ku, Nagoya, Aichi 4648601, Japan

来源：

FEBS LETTERS | 2003年 / 540卷 / 1-3期

基金：

日本学术振兴会;

关键词：

single-molecule polymerase chain reaction; in vitro coupled transcription/translation; combinatorial protein library; high-throughput screening;

D O I：

10.1016/S0014-5793(03)00251-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel entirely in vitro strategy for generation and screening of a combinatorial protein library in an array format has been developed and is experimentally demonstrated. The strategy exploits virtues of PCR and in vitro coupled transcription/translation. Our new approach provides high-throughput construction and screening of the in vitro protein library, and compatibility with various selection methods. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

引用

页码：147 / 150

页数：4

共 21 条

[11] Development of a microchamber array for picoliter PCR [J].

Nagai, H ;

Murakami, Y ;

Morita, Y ;

Yokoyama, K ;

Tamiya, E .

ANALYTICAL CHEMISTRY, 2001, 73 (05) :1043-1047

[12] Single-step single-molecule PCR of DNA with a homo-priming sequence using a single primer and hot-startable DNA polymerase [J].

Nakano, H ;

Kobayashi, K ;

Ohuchi, S ;

Sekiguchi, S ;

Yamane, T .

JOURNAL OF BIOSCIENCE AND BIOENGINEERING, 2000, 90 (04) :456-458

[13] In vitro method for the generation of protein libraries using PCR amplification of a single DNA molecule and coupled transcription/translation [J].

Ohuchi, S ;

Nakano, H ;

Yamane, T .

NUCLEIC ACIDS RESEARCH, 1998, 26 (19) :4339-4346

[14] Totally in vitro protein selection using mRNA-protein fusions and ribosome display [J].

Roberts, RW .

CURRENT OPINION IN CHEMICAL BIOLOGY, 1999, 3 (03) :268-273

[15] Phage-display technology - finding a needle in a vast molecular haystack [J].

Rodi, DJ ;

Makowski, L .

CURRENT OPINION IN BIOTECHNOLOGY, 1999, 10 (01) :87-93

[16] High-throughput, cloning-independent protein library construction by combining single-molecule DNA amplification with in vitro expression [J].

Rungpragayphan, S ;

Kawarasaki, Y ;

Imaeda, T ;

Kohda, K ;

Nakano, H ;

Yamane, T .

JOURNAL OF MOLECULAR BIOLOGY, 2002, 318 (02) :395-405

[17]

SCHOBER A, 1993, BIOTECHNIQUES, V15, P324

[18]

Tabuchi M, 2002, PROTEOMICS, V2, P430, DOI 10.1002/1615-9861(200204)2:4<430::AID-PROT430>3.0.CO

[19]

2-5

[20] Digital PCR [J].

Vogelstein, B ;

Kinzler, KW .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1999, 96 (16) :9236-9241

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