A versatile set of ligation-independent cloning vectors for functional studies in plants
| Autor: | Willy A. M. van den Berg, Hilda van Mourik, Che-Yang Liao, Dolf Weijers, Annemarie S. Lokerse, Cristina I. Llavata-Peris, Barbara Möller, Bert De Rybel |
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| Rok vydání: | 2011 |
| Předmět: |
0106 biological sciences
Physiology fluorescent proteins Genetic Vectors Molecular Sequence Data high-throughput cloning Arabidopsis Biochemie Plant Science Biology in-vitro 01 natural sciences Biochemistry 03 medical and health sciences Gene Expression Regulation Plant expression Genetics transcriptional regulation Vector (molecular biology) Cloning Molecular genes Promoter Regions Genetic Transcription factor Gene 030304 developmental biology Regulation of gene expression Cloning 0303 health sciences promoter EPS-1 Base Sequence Ligation-independent cloning Breakthrough Technologies root constructs arabidopsis Meristem initiation Function (biology) 010606 plant biology & botany |
| Zdroj: | Plant Physiology 156 (2011) Plant Physiology, 156, 1292-1299 |
| ISSN: | 1532-2548 0032-0889 |
| Popis: | With plant molecular biology in 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 ligation-independent 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. |
| Databáze: | OpenAIRE |
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