Search results for "Suppression subtractive hybridization"
showing 10 items of 11 documents
Sex versus parthenogenesis: A transcriptomic approach of photoperiod response in the model aphid Acyrthosiphon pisum (Hemiptera: Aphididae)
2007
International audience; Most aphids develop a cyclic parthenogenesis life-cycle. After several generations of viviparous parthenogenetic females, it follows a single annual generation of sexual individuals, usually in autumn, that mate and lay the sexual eggs. Shortening of photoperiod at the end of the summer is a key factor inducing the sexual response. With the survey here reported we aimed at identifying a collection of candidate genes to participate at some point in the cascade of events that lead to the sexual phenotypes. Following a suppression subtractive hybridization methodology (SSH) on the model aphid Acyrthosiphon pisum, we built and characterised two reciprocal cDNA libraries …
Functional genomics of arbuscular mycorrhiza : decoding the symbiotic cell programme
2004
More extensive insight into plant genes involved in the symbiotic programme of arbuscular mycorrhiza is presently being achieved by global approaches that aim to discover novel genes or subsets of genes that are essential to cell programmes in the different steps of plantfungal interactions. The strategy of functional genomics based on large-scale differential RNA expression analyses (differential-display reverse transcriptase - PCR), electronic Northerns, suppressive subtractive hybridization, DNA chips) is presented, with a focus on arbuscular mycorrhiza in Pisum sativum and Medicago truncatula. The most recent knowledge about gene networks that are modulated in roots during arbuscular …
Computational annotation of genes differentially expressed along olive fruit development
2009
Abstract Background Olea europaea L. is a traditional tree crop of the Mediterranean basin with a worldwide economical high impact. Differently from other fruit tree species, little is known about the physiological and molecular basis of the olive fruit development and a few sequences of genes and gene products are available for olive in public databases. This study deals with the identification of large sets of differentially expressed genes in developing olive fruits and the subsequent computational annotation by means of different software. Results mRNA from fruits of the cv. Leccino sampled at three different stages [i.e., initial fruit set (stage 1), completed pit hardening (stage 2) a…
LPS challenge regulates gene expression and tissue localization of a Ciona intestinalis gene through an alternative polyadenylation mechanism.
2013
A subtractive hybridization strategy for the identification of differentially expressed genes was performed between LPS-challenged and naive Ciona intestinalis. This strategy allowed the characterization of two transcripts (Ci8short and Ci8long) generated by the use of two Alternative Polyadenylation sites. The Ci8long transcript contains a protein domain with relevant homology to several components of the Receptor Transporting Protein (RTP) family not present in the Ci8short mRNA. By means of Real Time PCR and Northern Blot, the Ci8short and Ci8long transcripts showed a different pattern of gene expression with the Ci8short mRNA being strongly activated after LPS injection in the pharynx. …
Identification of DNA sequences specific for Vibrio vulnificus biotype 2 strains by suppression subtractive hybridization.
2005
ABSTRACT Vibrio vulnificus can be divided into three biotypes, and only biotype 2, which is further divided into serovars, contains eel-virulent strains. We compared the genomic DNA of a biotype 2 serovar E isolate (tester) with the genomic DNAs of three biotype 1 strains by suppression subtractive hybridization and then tested the distribution of the tester-specific DNA sequences in a wide collection of bacterial strains. In this way we identified three plasmid-borne DNA sequences that were specific for biotype 2 strains irrespective of the serovar and three chromosomal DNA sequences that were specific for serovar E biotype 2 strains. These sequences have potential for use in the diagnosis…
Effects of cadmium exposure on sea urchin development assessed by SSH and RT-qPCR: metallothionein genes and their differential induction
2012
In order to study the defense strategies activated by Paracentrotus lividus embryos in response to sub-lethal doses of CdCl2, we compared the induced transcripts to that of control embryos by suppression subtractive hybridization technique. We isolated five metallothionein (MT) cDNAs and other genes related to detoxification, to signaling pathway components, to oxidative, reductive and conjugative biotransformation, to RNA maturation and protein synthesis. RT-qPCR analysis revealed that two of the five P. lividus MT (PlMT7 and PlMT8) genes appeared to be constitutively expressed and upregulated following cadmium treatment, whereas the other three genes (PlMT4, PlMT5, PlMT6) are specifically…
Genes differentially expressed by Aspergillus carbonarius strains under ochratoxin A producing conditions
2010
Aspergillus carbonarius is an important ochratoxin A (OTA)-producing fungus that is responsible for toxin contamination of grapes and wine, coffee and cocoa. A suppression subtractive hybridization (SSH) approach was performed with two strains of A. carbonarius, antagonistic in their OTA-production ability, to identify genes whose expression is linked with the ability to produce OTA. BlastX analysis identified 109 differentially-expressed sequences putatively involved in the production of OTA, with significant similarities (Evalue < 10− 5) to sequences deposited in the NCBI non-redundant protein database. Of the 109 ESTs, 26% were involved in regulation processes, 15% corresponded to hypoth…
Differential display reverse transcription PCR reveals IL-1 induced gene expression patterns in human articular chondrocytes
1995
IL-1 exerts diverse effects on the metabolism of articular chondrocytes, including inhibition of proteoglycan synthesis and stimulation of matrix metallopro teinase synthesis. Therefore it is believed that IL-1 might play an important role in cartilage degradation in osteo- and rheumatoid arthritis. To improve our understanding of IL-1 induced effects on overall gene expression patterns of human articular chondrocytes, wc used a novel mRNA fingerprinting technique: Differential Display Reverse Transcription-PCR (DDRT-PCR) (Liang and Pardee 1992). The reported high sensitivity of this powerful technique promised to enable work with human articular cartilage, a tissue from which only small am…
A method to diagnose the carrier state of Vibrio vulnificus serovar E in eels: Development and field studies
2006
Abstract The pathogen Vibrio vulnificus serovar E (VSE) has been related to both human infections and to epizootics causing high mortality in brackish water eel farms. To control the spread of the eel vibriosis and prevent VSE transmission to humans we designed and tested a protocol to detect carriers, which involves isolating the pathogen. To identify the organs where VSE persists in survivors we infected eels with different degrees of immunity against the pathogen (non-immune [NI], immune [I, eels vaccinated 1 year before] and freshly vaccinated [V]) by bath challenge. Then, we followed the pathogen survival in selected external and internal organs for 72 h post-infection. VSE was isolate…
Isolation of a novel LPS-induced component of the ML superfamily in Ciona intestinalis
2015
ML superfamily represents a group of proteins playing important roles in lipid metabolism and innate immune response. In this study, we report the identification of the first component of the ML superfamily in the invertebrate Ciona intestinalis by means of a subtractive hybridization strategy. Sequence homology and phylogenetic analysis showed that this protein forms a specific clade with vertebrate components of the Niemann-Pick type C2 protein and, for this reason, it has been named Ci-NPC2. The putative Ci-NPC2 is a 150 amino acids long protein with a short signal peptide, seven cysteine residues, three putative lipid binding site and a three-dimensional model showing a characteristic b…