Cargando…
Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes
BACKGROUND: We have developed a functional genomics approach based on expression cloning in Xenopus oocytes to identify plant transporter function. We utilized the full-length cDNA databases to generate a normalized library consisting of 239 full-length Arabidopsis thaliana transporter cDNAs. The ge...
Autores principales: | , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2006
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1637106/ https://www.ncbi.nlm.nih.gov/pubmed/17069646 http://dx.doi.org/10.1186/1746-4811-2-17 |
_version_ | 1782130787042197504 |
---|---|
author | Nour-Eldin, Hussam H Nørholm, Morten HH Halkier, Barbara A |
author_facet | Nour-Eldin, Hussam H Nørholm, Morten HH Halkier, Barbara A |
author_sort | Nour-Eldin, Hussam H |
collection | PubMed |
description | BACKGROUND: We have developed a functional genomics approach based on expression cloning in Xenopus oocytes to identify plant transporter function. We utilized the full-length cDNA databases to generate a normalized library consisting of 239 full-length Arabidopsis thaliana transporter cDNAs. The genes were arranged into a 96-well format and optimized for expression in Xenopus oocytes by cloning each coding sequence into a Xenopus expression vector. RESULTS: Injection of 96 in vitro transcribed cRNAs from the library in pools of columns and rows into oocytes and subsequent screening for glucose uptake activity identified three glucose transporters. One of these, AtSTP13, had not previously been experimentally characterized. CONCLUSION: Expression of the library in Xenopus oocytes, combined with uptake assays, has great potential in assignment of plant transporter function and for identifying membrane transporters for the many plant metabolites where a transporter has not yet been identified. |
format | Text |
id | pubmed-1637106 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2006 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-16371062006-11-17 Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes Nour-Eldin, Hussam H Nørholm, Morten HH Halkier, Barbara A Plant Methods Methodology BACKGROUND: We have developed a functional genomics approach based on expression cloning in Xenopus oocytes to identify plant transporter function. We utilized the full-length cDNA databases to generate a normalized library consisting of 239 full-length Arabidopsis thaliana transporter cDNAs. The genes were arranged into a 96-well format and optimized for expression in Xenopus oocytes by cloning each coding sequence into a Xenopus expression vector. RESULTS: Injection of 96 in vitro transcribed cRNAs from the library in pools of columns and rows into oocytes and subsequent screening for glucose uptake activity identified three glucose transporters. One of these, AtSTP13, had not previously been experimentally characterized. CONCLUSION: Expression of the library in Xenopus oocytes, combined with uptake assays, has great potential in assignment of plant transporter function and for identifying membrane transporters for the many plant metabolites where a transporter has not yet been identified. BioMed Central 2006-10-27 /pmc/articles/PMC1637106/ /pubmed/17069646 http://dx.doi.org/10.1186/1746-4811-2-17 Text en Copyright © 2006 Nour-Eldin et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Nour-Eldin, Hussam H Nørholm, Morten HH Halkier, Barbara A Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes |
title | Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes |
title_full | Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes |
title_fullStr | Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes |
title_full_unstemmed | Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes |
title_short | Screening for plant transporter function by expressing a normalized Arabidopsis full-length cDNA library in Xenopus oocytes |
title_sort | screening for plant transporter function by expressing a normalized arabidopsis full-length cdna library in xenopus oocytes |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1637106/ https://www.ncbi.nlm.nih.gov/pubmed/17069646 http://dx.doi.org/10.1186/1746-4811-2-17 |
work_keys_str_mv | AT noureldinhussamh screeningforplanttransporterfunctionbyexpressinganormalizedarabidopsisfulllengthcdnalibraryinxenopusoocytes AT nørholmmortenhh screeningforplanttransporterfunctionbyexpressinganormalizedarabidopsisfulllengthcdnalibraryinxenopusoocytes AT halkierbarbaraa screeningforplanttransporterfunctionbyexpressinganormalizedarabidopsisfulllengthcdnalibraryinxenopusoocytes |