Bacteriophage lambda display of complex cDNA libraries: A new approach to functional genomics

被引：59

作者：

Santi, E ^{[1
]}

Capone, S ^{[1
]}

Mennuni, C ^{[1
]}

Lahm, A ^{[1
]}

Tramontano, A ^{[1
]}

Luzzago, A ^{[1
]}

Nicosia, A ^{[1
]}

机构：

[1] Ist Ric Biol Mol P Angeletti, I-00040 Pomezia, Italy

来源：

JOURNAL OF MOLECULAR BIOLOGY | 2000年 / 296卷 / 02期

关键词：

lambda phage; human brain cDNA library; mouse embryo cDNA library; GAP-43; EMX1-EMX2;

D O I：

10.1006/jmbi.1999.3471

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：

We describe the construction and characterization of two lambda surface displayed cDNA expression libraries derived from human brain and mouse embryo. cDNA inserts were obtained by tagged random-priming elongation of commercially available cDNA libraries and cloned into a novel lambda vector at the 3' end of the D capsid protein gene, which produced highly complex repertoires (1 x 10(8) and 2 x 10(7) phage). These libraries were affinity selected with a monoclonal antibody against the neural specific factor GAP-43 and with polyclonal antibodies that recognize the EMX1 and EMX2 homeoproteins. In both cases rapid identification of specific clones was achieved, which demonstrates the great potential of the lambda display system for generating affinity selectable cDNA libraries from complex genomes. (C) 2000 Academic Press.

引用

收藏

页码：497 / 508

页数：12

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