[20] Suppression subtractive hybridization: A versatile method for identifying differentially expressed genes

Diatchenko, Luda; Lukyanov, Sergey; Lau, Yun‐Fai Chris; Siebert, Paul D.

doi:10.1016/s0076-6879(99)03022-0

Cited by 360 publications

(246 citation statements)

References 16 publications

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“…The initiation and progression of leaf senescence is likely to be ß Blackwell Publishing Ltd, The Plant Journal, (2003), 36, 629±642 regulated primarily at the transcriptional level (He et al, 2001). The SSH method, which has proved to be a powerful tool for the ampli®cation of non-abundant transcripts in other systems (Diatchenko et al, 1999), allowed us to identify a number of new SAGs encoding potential regulators. In addition to the two reported senescence-related receptors, senescence-associated receptor kinase (SARK; Hajouj et al, 2000) and senescence-induced receptor kinase (SIRK; Robatzek and Somssich, 2002), we revealed in our screen a cDNA clone that represents a receptor-like protein kinase (At5g48380).…”

Section: Regulatory Genesmentioning

confidence: 99%

Large‐scale identification of leaf senescence‐associated genes

et al. 2003

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SummaryLeaf senescence is a form of programmed cell death, and is believed to involve preferential expression of a speci®c set of`senescence-associated genes' (SAGs). To decipher the molecular mechanisms and the predicted complex network of regulatory pathways involved in the senescence program, we have carried out a large-scale gene identi®cation study in a reference plant, Arabidopsis thaliana. Using suppression subtractive hybridization, we isolated approximately 800 cDNA clones representing SAGs expressed in senescing leaves. Differential expression was con®rmed by Northern blot analysis for 130 non-redundant genes. Over 70 of the identi®ed genes have not previously been shown to participate in the senescence process. SAGencoded proteins are likely to participate in macromolecule degradation, detoxi®cation of oxidative metabolites, induction of defense mechanisms, and signaling and regulatory events. Temporal expression pro®les of selected genes displayed several distinct patterns, from expression at a very early stage, to the terminal phase of the senescence syndrome. Expression of some of the novel SAGs, in response to age, leaf detachment, darkness, and ethylene and cytokinin treatment was compared. The large repertoire of SAGs identi®ed here provides global insights about regulatory, biochemical and cellular events occurring during leaf senescence.

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Section: Regulatory Genesmentioning

confidence: 99%

Large‐scale identification of leaf senescence‐associated genes

et al. 2003

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“…As with subtractive hybridization techniques [16], differential hybridization is advantageous in that it does not require a priori sequence information, but it is unique in that both commonly and specifically expressed mRNAs in multiple tissues or cell samples can be compared simultaneously. Thus, in the same experiment, genes expressed only in normal tissues (e.g., tumor suppressor genes), as well as genes that are tumor specific (e.g., tumor markers, activated oncogenes), can be identified [17].…”

Section: Identification Of the Genes Differentially Expressedmentioning

confidence: 99%

Downregulation of putative tumor suppressor gene TSC‐22 in human brain tumors

Shostak

Dmitrenko

Garifulin

et al. 2002

Journal of Surgical Oncology

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Background and Objectives: Our objective was to identify differentially expressed genes involved in the pathogenesis of glioblastoma multiforme (GBM). Methods: Screening of arrayed human fetal brain and human postnatal brain cDNA libraries was performed by differential hybridization with glioblastoma multiforme and human normal brain cDNAs. Results: Repeated differential hybridization of more than 100 cDNA clones selected by primary screening and analysis of RNA from adult normal brain and glial tumors showed 16 nucleotide sequences differentially expressed between normal brain and brain tumors. Among others, decreased content in astrocytic tumors was determined for TSC-22 mRNA corresponding to cDNA in the ICRFp507J1041 clone from human fetal brain cDNA library. Northern blot hybridization of RNA from different human brain tumors showed very low amounts of TSC-22 mRNA in most investigated samples of GBM, anaplastic astrocytoma, and some other tumors. Complete lack of expression of TSC-22 occurred in one sample of anaplastic astrocytoma, as well as in meningioma, brain sarcoma, sarcomatous meningioma, and oligodendroglioma. The differential expression of TSC-22 gene was confirmed by semiquantitative RT-PCR in 15 samples of astrocytomas WHO grade II-IV and three samples of normal brain. Conclusions: Significantly decreased levels of TSC-22 mRNA in human brain and salivary gland tumors and antiproliferative role of TSC-22 strongly suggest a tumor suppressor role for TSC-22.

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“…Second strand synthesis and RSA I digestion was according to standard procedures (Sambrook et al, 1989). Addition of linkers and all other procedures relevant to the SSH technique were derived either from Diatchenko et al (1999) or the supplier's recommendation (PCR-select cDNA subtraction, Clontech).…”

Section: Ssh Techniquementioning

confidence: 99%