Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.

Diatchenko, Luda; Lau, Yun‐Fai Chris; Campbell, Aaron P.; Chenchik, Alex; Moqadam, Fauzia; Huang, Betty; Lukyanov, Sergey; Lukyanov, Konstantin A.; Gurskaya, Nadya G.; Свердлов, Е. Д.; Siebert, Paul D.

doi:10.1073/pnas.93.12.6025

Cited by 2,768 publications

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“…Isolation of a cDNA which is upregulated in 208F cells transformed with v-fos and v-ras A cDNA library representing the mRNAs which are upregulated in FBR cells compared to 208F cells was prepared using the suppression subtractive hybridization method (Diatchenko et al, 1996), and 500 random clones were sequenced (Johnston et al, 1999, unpublished). The open reading frame (ORF) from one 400 bp clone, designated krp1, showed 31% amino acid sequence identity to a Drosophila kelch protein, which is involved in crosslinking actin ®laments in the ring canals during development (Xue and Cooley, 1993;Robinson and Cooley, 1997).…”

Section: Resultsmentioning

confidence: 99%

“…Examples of proteins expressed are type 1 collagenase and stromelysin 1, both of which are proteases that are involved in the degradation of the ECM (Kerr et al, 1988;Schonthal et al, 1988). In order to identify more AP-1 regulated genes that are needed for FBR invasion, we have used di erential screening and subtractive suppression hybridization (Diatchenko et al, 1996;Hennigan et al, 1994;Johnston et al, 1999, unpublished). These searches have revealed genes which encode proteins that have previously been implicated in invasion, as well as numerous novel genes that may play a role in transformation.…”

Section: Introductionmentioning

confidence: 99%

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Krp1, a novel kelch related protein that is involved in pseudopod elongation in transformed cells

et al. 2000

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Krp1, a novel kelch related protein that is involved in pseudopod elongation in transformed cells

et al. 2000

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“…cDNA subtraction was done using Clontech's Suppression Subtractive Hybridization (SSH) kit following the manufacturer's protocol (Diatchenko et al 1996) with 33 and 21 cycles for primary and secondary PCR respectively. Forward subtraction was done using cDNA synthesised from seeds categorized as 'tester' from mid-to-late maturation stage (JC3) and as 'driver' from early-to-mid maturation (JC2) stage (Fig.…”

Section: Ssh Library Preparation and Est Sequencingmentioning

confidence: 99%

“…Here we report a suppression subtractive hybridization (SSH) based approach to identify differentially expressing genes in different maturation stages of J. curcas seeds that correspond to active oil metabolism. SSH is a powerful technique that enriches differentially expressing genes (Diatchenko et al 1996) and has been used to study seed development related genes in Phaseolus vulgaris (Abid et al 2012). In Brassica napus, differentially expressing genes in seeds of two near-isogenic lines having different oil content were investigated using SSH.…”

Section: Introductionmentioning

confidence: 99%

Differential expression analysis of transcripts related to oil metabolism in maturing seeds of Jatropha curcas L.

Chandran

Sankararamasubramanian

Kumar

et al. 2014

Physiol Mol Biol Plants

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Jatropha curcas has been widely studied at the molecular level due to its potential as an alternative source of fuel. Many of the reports till date on this plant have focussed mainly on genes contributing to the accumulation of oil in its seeds. A suppression subtractive hybridization strategy was employed to identify genes which are differentially expressed in the mid maturation stage of J. curcas seeds. Random expressed sequence tag sequencing of the cDNA subtraction library resulted in 385 contigs and 1,428 singletons, with 591 expressed sequence tags mapping for enzymes having catalytic roles in various metabolic pathways. Differences in transcript levels in early and mid-to-late maturation stages of seeds were also investigated using sequence information obtained from the cDNA subtraction library. Seven out of 12 transcripts having putative roles in central carbon metabolism were up regulated in early seed maturation stage while lipid metabolism related transcripts were detected at higher levels in the later stage of seed maturation. Interestingly, 4 of the transcripts revealed putative alternative splice variants that were specifically present or up regulated in the early or late maturation stage of the seeds. Transcript expression patterns from the current study using maturing seeds of J. curcas reveal a subtle balancing of oil accumulation and utilization, which may be influenced by their energy requirements.

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“…In this study, we have performed Representational Difference Analysis (RDA) (Diatchenko et al, 1996) in order to identify novel genes expressed in normal epithelium but absent from SCC. This study has allowed isolation of a differentially expressed novel gene named DESC1.…”

mentioning

confidence: 99%

Differential expression of a novel serine protease homologue in squamous cell carcinoma of the head and neck

Lang

Schuller

2001

Br J Cancer

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Summary Differential gene expression between squamous cell carcinoma of the head and neck and matched normal tissue was studied by utilizing Representational Difference Analysis. Using this methodology, a novel gene, DESC1 was isolated. DESC1 possesses strong identity to the serine protease super-family. Comparison of DESC1 expression between primary squamous cell carcinoma and matched normal tissue shows that the level of DESC1 expression is reduced or absent in 11/12 SCC tissue specimens when compared to specimens of matched normal tissue. Tissue-specific expression studies further show that DESC1 expression can only be detected in tissues derived from the head and neck, and in skin, prostate and testes. Cell line studies demonstrate that DESC1 expression is epithelial-specific. Chromosomal localization studies indicate that DESC1 is located on the long arm of chromosome 4 at position q12-13.

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Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.

Cited by 2,768 publications

References 19 publications

Krp1, a novel kelch related protein that is involved in pseudopod elongation in transformed cells

Krp1, a novel kelch related protein that is involved in pseudopod elongation in transformed cells

Differential expression analysis of transcripts related to oil metabolism in maturing seeds of Jatropha curcas L.

Differential expression of a novel serine protease homologue in squamous cell carcinoma of the head and neck

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