A semi-automated high-throughput approach to the generation of transposon insertion mutants in the nematode Caenorhabditis elegans

被引：21

作者：

Duverger, Yohann

Belougne, Jerome

Scaglione, Sarah

Brandli, Dominique

Beclin, Christophe

Ewbank, Jonathan J.

机构：

[1] Univ Mediterranee, Ctr Immunol, F-13288 Marseille 9, France

[2] INSERM, U631, F-13288 Marseille, France

[3] CNRS, UMR6102, F-13288 Marseille, France

[4] CNRS, Inst Dev Biol, Marseille, France

来源：

NUCLEIC ACIDS RESEARCH | 2007年 / 35卷 / 02期

关键词：

D O I：

10.1093/nar/gkl1046

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：

The generation of a large collection of defined transposon insertion mutants is of general interest to the Caenorhabditis elegans research community and has been supported by the European Union. We describe here a semi-automated high-throughput method for mutant production and screening, using the heterologous transposon Mos1. The procedure allows routine culture of several thousand independent nematode strains in parallel for multiple generations before stereotyped molecular analyses. Using this method, we have already generated > 17 500 individual strains carrying Mos1 insertions. It could be easily adapted to forward and reverse genetic screens and may influence researchers faced with making a choice of model organism.

引用

收藏

页数：8

相关论文

共 24 条

[1] Targeted gene alteration in Caenorhabditis elegans by gene conversion [J].

Barrett, PL ;

Fleming, JT ;

Göbel, V .

NATURE GENETICS, 2004, 36 (11) :1231-1237

[2] Homologous gene targeting in Caenorhabditis elegans by biolistic transformation -: art. no. e40 [J].

Berezikov, E ;

Bargmann, CI ;

Plasterk, RHA .

NUCLEIC ACIDS RESEARCH, 2004, 32 (04) :e40

[3] Mobilization of a Drosophila transposon in the Caenorhabditis elegans germ line [J].

Bessereau, JL ;

Wright, A ;

Williams, DC ;

Schuske, K ;

Davis, MW ;

Jorgensen, EM .

NATURE, 2001, 413 (6851) :70-74

[4] TLR-independent control of innate immunity in Caenorhabditis elegans by the TIR domain adaptor protein TIR-1, an ortholog of human SARM [J].

Couillault, C ;

Pujol, N ;

Reboul, J ;

Sabatier, L ;

Guichou, JF ;

Kohara, Y ;

Ewbank, JJ .

NATURE IMMUNOLOGY, 2004, 5 (05) :488-494

[5] Improved detection of small deletions in complex pools of DNA [J].

Edgley, M ;

D'Souza, A ;

Moulder, G ;

McKay, S ;

Shen, B ;

Gilchrist, E ;

Moerman, D ;

Barstead, R .

NUCLEIC ACIDS RESEARCH, 2002, 30 (12) :e52

[6] Functional genomic analysis of C-elegans chromosome I by systematic RNA interference [J].

Fraser, AG ;

Kamath, RS ;

Zipperlen, P ;

Martinez-Campos, M ;

Sohrmann, M ;

Ahringer, J .

NATURE, 2000, 408 (6810) :325-330

[7] Characterization of mutations induced by ethyl methanesulfonate, UV, and trimethylpsoralen in the nematode Caenorhabditis elegans [J].

Gengyo-Ando, K ;

Mitani, S .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2000, 269 (01) :64-69

[8] Functional genomic analysis of cell division in C-elegans using RNAi of genes on chromosome III [J].

Gönczy, P ;

Echeverri, C ;

Oegema, K ;

Coulson, A ;

Jones, SJM ;

Copley, RR ;

Duperon, J ;

Oegema, J ;

Brehm, M ;

Cassin, E ;

Hannak, E ;

Kirkham, M ;

Pichler, S ;

Flohrs, K ;

Goessen, A ;

Leidel, S ;

Alleaume, AM ;

Martin, C ;

Özlü, N ;

Bork, P ;

Hyman, AA .

NATURE, 2000, 408 (6810) :331-336

[9] Mos as a tool for genome-wide insertional mutagenesis in Caenorhabditis elegans:: results of a pilot study -: art. no. e117 [J].

Granger, L ;

Martin, E ;

Ségalat, L .

NUCLEIC ACIDS RESEARCH, 2004, 32 (14) :e117

[10] Isolation of C-elegans deletion mutants following ENU mutagenesis and thermostable restriction enzyme PCR screening [J].

Huang, CG ;

Agre, P ;

Strange, K ;

Lamitina, T .

MOLECULAR BIOTECHNOLOGY, 2006, 32 (01) :83-86