• 한국어병학회지
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• 제19권3호
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• pp.189-195
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• 2006

A subtracted cDNA library of a marine scuticociliate, Philasterides dicentrarchii, in response to 17β-estradiol exposure was constructed using suppression subtractive hybridization (SSH). As a result of SSH, 275 clones were isolated, and among them, only glutathione peroxidase (GPX) gene was isolated as an antioxidative enzyme responding to 17β-estradiol. The semi-quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) analysis revealed that the transcription of GPX gene of P. dicentrarchii was clearly increased by exposure to 17β-estradiol. The GPX transcription was also clearly increased by exposure to xenoestrogens such as bisphenol A (BPA) and genistein.

• 한국자원식물학회지
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• 제19권3호
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• pp.440-446
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• 2006

To investigate flowering related genes in winter-type oilseed rape (Brassica napus L. cv. Tammi), differentially expressed genes were isolated from leaves of the plant after low temperature treatment which is requirements for floral induction. As a result of suppression subtractive hybridization (SSH), 288 clones were randomly selected from SSH library. Using reverse Northern blot analysis, 150 of 288 clones were identified to be differentially expressed. Out of these 150 clones, 45 clones showed very high identities with the known genes. Four clones showed very high identities over 90% with metallothionein-like gene that is related to flowering-induced genes. Of these 4 clones, the cDNA clone, rfs-13, revealed high identity with meotallothionein-like protein in Arabidopsis thaliana (98%) and Brassica compestris (89%). Furthermore, gene expressed in immature flower stages was confirmed by Northern blot analysis.

• 한국생명과학회:학술대회논문집
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• 한국생명과학회 2001년도 10주년 기념 국제학술 심포지움 및 추계학술대회
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• pp.64-66
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• 2001

• 한국육종학회지
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• 제40권3호
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• pp.234-242
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• 2008

To identify low temperature-induced genes of wheat chromosome substitution line 5D, suppression subtractive hybridization (SSH) was performed with mRNAs from leaf samples that treated with low temperature ($4^{\circ}C$). A cDNA library was constructed using mRNA isolated from wheat chromosome substitution line 5D leaves treated with low temperature ($4^{\circ}C$). The nucleotide and deduced amino acid sequences of the putative gene products were compared. wfr-9 and wfr-32 showed identity over 90% related to vernalization gene. Other two genes, wfr-77 and wfr-83 which is related to freezing-resistant gene have also identity over 90%. This result suggest that those genes may be transcribed into antifreeze proteins which are accumulated within leaf apoplasts, when wheat chromosome substitution line 5D is acclimated during low temperature treatment.

• 한국육종학회지
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• 제41권2호
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• pp.73-83
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• 2009

To investigate genes related to vernalization and cold- resistance in barley (Hordeum vulgare L. cv. Nagaoka), differentially expressed genes were identified from cold-resistant barley leaves with suppression subtractive hybridization (SSH) and Northern blot analyses. The nucleotide and the deduced amino acid sequences of the putative gene products were compared. The bvrn-7 showed high homology(84%) with gene related to vernalization, and the bvrn-3, bvrn-12, bvrn-28, bvrn-29 and bvrn-36 related to cold-resistant genes had high identity of 88~98% with low temperature-induced genes. The results indicate that the 6 genes were closely related to vernalization and cold-resistance during low temperature treatment.

• Asian-Australasian Journal of Animal Sciences
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• 제23권1호
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• pp.17-24
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• 2010

In the antral follicle phase, several layers of cumulus cells surround the oocyte and play an important support and regulation role in oocyte development and maturation via intercellular communications and interactions between oocytes and cumulus cells. However, information on stage specific gene expression in swine during the phase is not well understood. To investigate the function of cumulus cells during in vitro maturation of porcine oocytes and gene expression, suppression subtractive hybridization (SSH) was performed to screen genes that were differentially expressed between cumulus-oocyte complexes (COCs) and naked oocytes (NOs). Utilizing mRNAs from in vitro maturation oocytes, a SSH cDNA library from COCs as the tester and NOs as the driver was constructed. The SSH cDNA library was then screened using dot blot analysis. Results showed that a total of 70 clones randomly selected from the library were differentially expressed. Among these, 41 exhibited high homology to known genes and 11 were novel expressed sequences tags (ESTs). Four differentially expressed genes, including bfgf, sprouty 2, egr and btc, were further studied by real time quantitative PCR; results confirmed an increased expression of respective mRNA in COCs compared with NOs, which suggests that these factors may play an important role in oocyte development and maturation.

• 대한의생명과학회지
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• 제10권3호
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• pp.195-202
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• 2004

Lung cancer is a leading cause of cancer death worldwide; however, despite major advances in cancer treatment during the past two decades, the prognostic outcome of lung cancer patients has improved only minimally. This is largely due to the inadequacy of the traditional screening approach of diagnosis in lung cancer, which detects only well­established overt cancers and fails to identify precursor lesions in premalignant conditions of the bronchial tree. In recent years this situation has fundamentally changed with the identification of molecular abnormalities characteristic of premalignant changes; these concern tumour suppressor genes, loss of heterozygosity at crucial sites and activation of oncogenes. Basic knowledge at the molecular level has extremely important clinical implications with regard to early diagnosis, risk assessment and prevention, and therapeutic targets. In this study we used a 'cap-finder' subtractive hybridization method, 'long distance' polymerase chain reaction (PCR), streptavidin magnetic beads mediated subtraction, and spin column chromatography to detect differential expression genes of human small cell lung carcinoma. We have now isolated ninety two genes that expressed differentially in the human small cell lung carcinoma cells and analyzed of 12 clones with sequencing, nine cDNAs include tapasin (NGS-17) mRNA, BC200 alpha scRNA, chromosome 12q24 PAC RPCI3-462E2, protein phosphatase 1 (PPPICA), translocation protein 1 (TLOC1), ribosomal protein S24 (RPS24) mRNA, protein phosphatase (PPEF2), cathepsin Z, MDM2 gene and three novel genes. They may be oncogenesis­related proteins.

• 대한의생명과학회지
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• 제8권3호
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• pp.195-202
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• 2002

Osteoarthritis (OA), the most common form of arthritis, involves the destabilization of the normal balance between the degradation and the synthesis of articular cartilage and subchondral bone within a joint. As articular cartilage degrades over time, its smooth surface roughens and bone-against-bone contact ensues, producing the inflammation response symptomatic of this 'wear and tear' disease. Although a variety of genetic, developmental, metabolic, and traumatic factors may initiate the development of osteoarthritis, its symptoms (joint pain, stiffness, and curtailed function) typically evolve slowly, and patients experience periods of relative calm alternation with episodes of inflammation and pain. Rheumatoid arthritis (RA), an autoimmune disease of unknown etiology characterized by chronic synovitis and cartilage destruction, affect 1% of the total population. Cartilage is a specialized connective tissue in which the chondrocytes occupy only 5% of the volume. Cartilage is particularly rich in extracellular matrix, with matrix making up 90% of the dry weight of the tissue chondrocytes have cell processes that extend a short distance into the matrix, but do not touch other cells thus in cartilage, cell-matrix interactions are essential for the maintenance of the extracellular matrix. In this study, subtractive hybridization method was utilized to detect genes differentially expressed in chondrocytes treated with methylprednisolone. We have isolated 57 genes that expressed differentially in the chondreocytes by methylprednisolone. 13 clones of them were analyzed with sequencing and their homologies were searched. 8 cDNAS included KIAA 0368, upregulated during skeletal muscle growth 5 (usmg 5), ribosomal protein S 18 (RPS 18), skeletal muscle ryanodine receptor, radial spoke protein 3 (RSP 3), ribosomal protein QM, ribosomal protein L37a (RPL37A), cytochrome coxidase subunit VIII (COX8).

• Journal of Plant Biotechnology
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• 제29권2호
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• pp.79-84
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• 2002

Suppression subtractive hybridization (SSH)를 통해 상처에 의해 발현이 유도되는 cDNA들을 분리하였고, 그 중 하나인 gmwi33은 $\beta$-amyrin synthase 유전자들과 높은 유사성을 보였다. gmwi33의 전장 cDNA인 GmAMS1은 2416 bp 길이에 739개 아미노산으로 구성된 긴 open reading frame(ORF)를 포함하고 있었다. GmAMS1 단백질은 감초의 $\beta$-amyrin synthase인 GgbAS와 89%, 완두의 OSCPSY와 86%의 유사성을 보였다. 암조건 하에 5일간 기른 콩나물에서, GmAMS1는 빛을 쪼여주었을 때 가장 강하게 발현되었고 methyl jasmonate 처리와 저온처리 시에도 발현이 유도된 반면, UV-B나 elicitor를 처리하였을 때는 발현이 유도되지 않았다. 이러한 GmAMS1의 발현양상은 사포닌의 활성산소 제거기능과 밀접한 연관이 있을 것으로 추측된다.

• Radiation Oncology Journal
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• 제23권1호
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• pp.43-50
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• 2005

목적 : 자경경부암세포에서 polymeric chain reaction (PCR)원리를 이용한 suppression subtractive hybridization (SSH) 방법으로 방사선조사 시 차등 발현되는 유전자를 동정하고자 하였다. 대상 및 방법 : 자궁경부암세포주인 HeLa세포주에 방사선조사 전과 후 총 RNA와 poly $(A)^+$ mRNA를 분리였다. SSH방법으로 forward 및 reverse-subtracted cDNA libraries를 만들었다. 차등 발현된 유전자를 screening하기 위해 reverse Northern blotting (dot blot analysis)을 이용하여 각각의 library에서 88개의 클론을 선택하였고 Nothern blotting으로 확인 후 sequencing하였다. 결과 : screening상 176개 클론 중 forward-subtracted library에서 10개의 유전자가 reverse-subtracted library에서 9개의 유전자가 동정되었다. forward-subtracted library로부터 3개의 유전자가 Northern blotting에 의하여 확인되었고 이중 telomerase catalytic subunit and sodium channel-like protein 유전자와 1개의 ESTs (expressed sequence tags) 유전자가 방사선선량에 따라 증가하쳐다. 결론 : 본 연구를 통해 자궁경부암세포주에서 방사선에 의해 유도되는 유전자를 SSH 방법을 통해 동정할 수 있었다. 그러나 이러한 유전자가 어떤 생물학적인 기능을 갖고 있는지에 대한 계속적인 연구가 필요하다