A versatile set of ligation-independent cloning vectors for functional studies in plants

Bert D. de Rybel; Willy van Den Berg; Annemarie Lokerse; Che Yang Liao; Hilda Van Mourik; Barbara Möller; Cristina Llavata Peris; Dolf Weijers

doi:10.1104/pp.111.177337

A versatile set of ligation-independent cloning vectors for functional studies in plants

Bert D. de Rybel, Willy van Den Berg, Annemarie Lokerse, Che Yang Liao, Hilda Van Mourik, Barbara Möller, Cristina Llavata Peris, Dolf Weijers^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

With plant molecular biology entering the omics era, there is a need for simple cloning strategies that allow high throughput to systematically study the expression and function of large numbers of genes. Such strategies would facilitate the analysis of gene (sub)families and/or sets of coexpressed genes identified by transcriptomics. Here, we provide a set of 34 ligationindependent cloning (LIC) binary vectors for expression analysis, protein localization studies, and misexpression that will be made freely available. This set of plant LIC vectors offers a fast alternative to standard cloning strategies involving ligase or recombination enzyme technology. We demonstrate the use of this strategy and our new vectors by analyzing the expression domains of genes belonging to two subclades of the basic helix-loop-helix transcription factor family. We show that neither the closest homologs of TARGET OF MONOPTEROS7 (TMO7/ATBS1) nor the members of the ATBS1 INTERACTING FACTOR subclade of putative TMO7 interactors are expressed in the embryo and that there is very limited coexpression in the primary root meristem. This suggests that these basic helix-loop-helix transcription factors are most likely not involved in TMO7-dependent root meristem initiation.

Original language	English
Pages (from-to)	1292-1299
Number of pages	8
Journal	Plant Physiology
Volume	156
Issue number	3
DOIs	https://doi.org/10.1104/pp.111.177337
Publication status	Published - Jul 2011
Externally published	Yes

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title = "A versatile set of ligation-independent cloning vectors for functional studies in plants",

abstract = "With plant molecular biology entering the omics era, there is a need for simple cloning strategies that allow high throughput to systematically study the expression and function of large numbers of genes. Such strategies would facilitate the analysis of gene (sub)families and/or sets of coexpressed genes identified by transcriptomics. Here, we provide a set of 34 ligationindependent cloning (LIC) binary vectors for expression analysis, protein localization studies, and misexpression that will be made freely available. This set of plant LIC vectors offers a fast alternative to standard cloning strategies involving ligase or recombination enzyme technology. We demonstrate the use of this strategy and our new vectors by analyzing the expression domains of genes belonging to two subclades of the basic helix-loop-helix transcription factor family. We show that neither the closest homologs of TARGET OF MONOPTEROS7 (TMO7/ATBS1) nor the members of the ATBS1 INTERACTING FACTOR subclade of putative TMO7 interactors are expressed in the embryo and that there is very limited coexpression in the primary root meristem. This suggests that these basic helix-loop-helix transcription factors are most likely not involved in TMO7-dependent root meristem initiation.",

author = "{de Rybel}, {Bert D.} and {van Den Berg}, Willy and Annemarie Lokerse and Liao, {Che Yang} and {Van Mourik}, Hilda and Barbara M{\"o}ller and Peris, {Cristina Llavata} and Dolf Weijers",

year = "2011",

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doi = "10.1104/pp.111.177337",

language = "English",

volume = "156",

pages = "1292--1299",

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AU - Lokerse, Annemarie

AU - Liao, Che Yang

AU - Van Mourik, Hilda

AU - Möller, Barbara

AU - Peris, Cristina Llavata

AU - Weijers, Dolf

PY - 2011/7

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A versatile set of ligation-independent cloning vectors for functional studies in plants

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