A specific member of the Cab multigene family can be efficiently targeted and disrupted in the moss Physcomitrella patens.

Détails

ID Serval
serval:BIB_D7C8C32864E9
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
A specific member of the Cab multigene family can be efficiently targeted and disrupted in the moss Physcomitrella patens.
Périodique
Molecular & Amp; General Genetics
Auteur(s)
Hofmann A.H., Codón A.C., Ivascu C., Russo V.E., Knight C., Cove D., Schaefer D.G., Chakhparonian M., Zrÿd J.P.
ISSN
0026-8925
Statut éditorial
Publié
Date de publication
02/1999
Peer-reviewed
Oui
Volume
261
Numéro
1
Pages
92-99
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't - Publication Status: ppublish
Résumé
The analysis of phenotypic change resulting from gene disruption following homologous recombination provides a powerful technique for the study of gene function. This technique has so far been difficult to apply to plants because the frequency of gene disruption following transformation with constructs containing DNA homologous to genomic sequences is low (0.01 to 0.1%). It has recently been shown that high rates of gene disruption (up to 90%) can be achieved in the moss Physcomitrella patens using genomic sequences of unknown function. We have used this system to examine the specificity of gene disruption in Physcomitrella using a member of the Cab multigene family. We have employed the previously characterised Cab gene ZLAB1 and have isolated segments of 13 other closely related members of the Cab gene family. In the 199-bp stretch sequenced, the 13 new members of the Cab family show an average of 8.5% divergence from the DNA sequence of ZLAB1. We observed 304 silent substitutions and 16 substitutions that lead to a change in the amino acid sequence of the protein. We cloned 1029 bp of the coding region of ZLAB1 (including 177 of the 199 bp with high homology to the 13 new Cab genes) into a vector containing a selectable hygromycin resistance marker, and used this construct to transform P. patens. In three of nine stable transformants tested, the construct had inserted in, and disrupted, the ZLAB1 gene. There was no discernible phenotype associated with the disruption. We have therefore shown that gene disruption is reproducible in P. patens and that the requirement for sequence homology appears to be stringent, therefore allowing the role of individual members of a gene family to be analysed in land plants for the first time.
Mots-clé
Amino Acid Sequence, Base Sequence, Blotting, Southern, Bryopsida/genetics, DNA, Plant/chemistry, DNA, Plant/genetics, Genes, Plant/genetics, Molecular Sequence Data, Multigene Family/genetics, Mutagenesis, Site-Directed, Polymerase Chain Reaction, Sequence Analysis, DNA, Sequence Homology, Nucleic Acid, Transformation, Genetic
Pubmed
Web of science
Création de la notice
13/03/2008 19:29
Dernière modification de la notice
20/08/2019 16:57
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