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Eur. J. Phycol. (2001), 36: 359366. Printed in the United Kingdom 359 Suppression subtractive hybridization for studying gene
 

Summary: Eur. J. Phycol. (2001), 36: 359366. Printed in the United Kingdom 359
Suppression subtractive hybridization for studying gene
expression during aerial exposure and desiccation in fucoid
algae
GARETH PEARSON, ESTER A. SERRAM O AND M. LEONOR CANCELA
Centro de CieV ncias do Mar, Unidade de CieV ncias e Tecnologias dos Recursos AquaT ticos (UCTRA), Universidade do Algarve,
Gambelas, 8000-810 Faro, Portugal
(Received 15 October 2000; accepted 10 May 2001)
Gene expression in Fucus vesiculosus L. from the Ria Formosa, Faro, Portugal was investigated by screening a cDNA
library generated by suppression subtractive hybridization (SSH) for algae undergoing mild desiccation stress (6070%
tissue water content). The subtractive library was constructed from small amounts (1 g) of total RNA using PCR-based
techniques. Screening by reverse Northern analysis (cDNA Southerns) revealed that between 60% and 70% of clones
randomly selected from the library were differentially regulated in desiccated and hydrated algae. Most genes could not be
directly identified based on sequence homology with known gene sequences. However, several cDNAs for chloroplast-
encoded transcripts were identified and shown to be up-regulated or differentially regulated in desiccated algae relative to
hydrated controls. These included partial sequences for ribulose-l,5-bisphosphate carboxylase\oxygenase (Rubisco;
rbcL\rbcS), chloroplast coupling factor ATPase (atpH\atpI) and a photosystem I P700 chlorophyll a binding protein
(psaA). The usefulness of reverse Northern analysis was confirmed with conventional Northern analysis of transcript
abundance for Rubisco, which varied rapidly in response to light and the hydration status of the algae. These results show
that SSH is a useful technique for analysing the responses of gene expression to environmental stress, and as a starting

  

Source: Alberto, Filipe - Centro de Ciencias do Mar, Universidade do Algarve

 

Collections: Environmental Sciences and Ecology; Biology and Medicine