• IMMUNE NETWORK
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• 제4권4호
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• pp.237-243
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• 2004

Background: The normal functions of the cell cycle inhibitor p16INK4a are frequently inactivated in many human cancers. Over 80% of hepatocellular carcinoma (HCC) cases lack a functional p16/Rb pathway. p16/Rb pathway, as well as p53 pathway, is considered as one of key components of tumor suppression. Methods: To study the roles of p16INK4a in HCC, a stable cell line expressing exogenous p16 was generated from SNU-449 hepatocellular carcinoma cells lacking endogenous p16, and suppression subtractive hybridization (SSH) was performed in parallel with the control cells. Results: 1) SSH identifies fibronectin (FN1), crystallin ${\alpha}B$ (CRYAB), Rac1, WASP, RhoGEF, and CCT3 as differentially-expressed genes. 2) Among the selected genes, the up-regulation of FN1 and CRYAB was confirmed by Northern blot, RT-PCR and by proteomic methods. Conclusion: These genes are likely to be associated with the induction of stress fiber and stabilization of cytoskeleton. Further studies are required to clarify the possible role of p16 in the signal transduction pathway.

• Journal of Life Science
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• 제11권1호
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• pp.52-56
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• 2001

The present study intends to characterize the DNA damage-inducible responses in eukaryotic cells. The fission yeast, S. pombe, which displays efficient DNA repair systems, was used in this study as a model system for higher eukaryotes. To study UV-inducible responses in S. pombe, five UV-inducible cDNA clones were isolated from S. pombe by using subtration hybridization method. To investigate the expression of isolated genes, the cellular levels of the transcripts of these genes were determined by Northern blot analysis after UV-irradiation. The transcripts of isolated gene (UV130) increased rapidly and reached maximum accumulation after UV-irradiation. Compared to the message levels of control, the levels of maximal increase were approximately 5 fold to UV-irradiation. In order to investigation whether the increase of UV130 transcripts was a specific results of UV-irradiation, UV130 transcript levels were examined after treating the cells to Methylmethane sulfonate (MMS). The transcripts of UV130 were not induced by treatment of 0.25% MMS. These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of these genes. To characterize the structure of UV130 gene, nucleotide sequences were analyzed. The nucleotide sequence of 1,340 nucleotide excluding poly(A) tail contains one open reading frame, which encodes a protein of 270 amino acids. The predicted amino acid sequences of UV130 do not exhibit any significant similarity to ther known sequences in the database.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권2호
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• pp.223-228
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• 2008

A novel beetle antimicrobial protein from stimulated Copris tripartitus and the corresponding gene were isolated in parallel through differential display-PCR and expression in Escherichia coli. To find cDNA clones responsible for bacteria resistance, the suppression subtractive hybridization and GeneFishing differentially expressed genes system were employed in the dung beetle, Copris tripartitus immunized with lipopolysaccaride. One cDNA clone from eight subtracted clones was selected through dot blot analysis and confirmed by northern blot analysis. The 516-bp, selected cDNA clone was determined by 5' and 3' rapid amplication of cDNA ends and cloned into the GST fusion expression vector pGEX-4T-1 for expression of the protein. The expressed protein was predicted 14.7 kDa and inhibited the growth of gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa. These results implied that the expressed protein is related to immune defense mechanism against microorganism.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권1호
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• pp.11-18
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• 2008

Human bone marrow mesenchymal stem cells are thought to be multipotent cells, which are present in adult marrow, that can replicate as undifferentiated cells and that have the potential to differentiate to lineages of mesenchymal tissues, including bone, cartilage, fat, tenden, muscle, and marrow stroma. Cells that have the characteristics of human mesenchymal stem cells could be isolated from marrow aspirates of human and animals. This study was designed to identify and characterize genes specifically expressed by osteogenic supplements -treated cells by suppression subtractive hybridization(SSH) method. The results were as follows: 1. 2 genes were upregulated genes in osteogenic diffeentiation of hMSCs, which is further proved by Northern blot analysis. 2. IGFBP-2 has been identified playing an important role in bone formation. 3. HF1 was also upregulated during osteogenic differentiation, but its role in bone formation is not clear yet.

• 생명과학회지
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• 제26권10호
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• pp.1218-1223
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• 2016

전복은 전복과 전복속에 속하는 복족류 연체동물로, 전 세계적으로 어업에 의한 식용전복의 공급은 감소하고, 양식에 의한 전복생산은 증가하고 있다. 한국, 일본, 중국을 포함해 전 세계적으로 전복은 중요한 해양 식량자원이며, 양식 산업에 있어서도 그 비중이 증가되고 있긴 하나, 그 느린 성장 속도가 가장 큰 문제점 중의 하나로 여겨진다. 선별, 종내교잡, 종간교잡, 배수체 등과 같은 여러 진보된 기술의 도입을 통하여 전복의 주요 양식 품종을 개발하기 위한 연구가 시도되고 있다. 양식용 전복품종개발을 위하여, 한국 해안에 서식하고 있는 전복은 6종(Haliotis discus hannai, 북방전복; Haliotis discus discus, 둥근전복; Haliotis makada, 왕전복; Haliotis gigantea, 말전복; Haliotis diversicolor diversicolor, 마대오분자기; Haliotis diversicolor supertexta, 오분자기) 간의 교배를 통한 교배종 개발이 유용한 기술 중의 하나가 될 수 있으며, 한국, 중국, 그리고 일본에 서식하는 종 간의 교배종 연구가 활발히 진행 되어 오고 있다. 유용한 잠재적 형질을 가진 한국 산 전복들의 계통학적 관계와 이들의 종 판별에 대해서 살펴볼 것이며, 이들 종들로부터 개발된 분자 마커의 현황과, 성공적인 양식용 교잡종 개발에 있어서 고려해야 할 유전적 사항들에 대하여 알아보고자 한다.

• Journal of Forest and Environmental Science
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• 제33권3호
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• pp.197-201
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• 2017

Populus. caspica L. is an Iranian indigenous poplar species which naturally distributed in the northern part of country. Unfortunately, overuse has removed many of the stems of better form, so that natural stands now usually appear small and crook. Therefore genetic variation for selection of new superior clone of this species is needed. Conventional hybridization system is currently used to induce genetic variation in poplar species but incompatibility barriers have been observed between them. In vitro ovule embryo culture was used to overcome incompatibility obstacle for interspecific hybridization between Populus caspica L. with Populus deltoids L.62/75. Female flowers of Populus caspica L. have artificially been pollinated with pollen grain of P. deltoides 62/75 in one direction using twig and pot crossing system. Ovaries at different ages (7, 14 and 21 days after pollination) were disinfected through 70% ethanol for 1 minute, 5% of sodium-hypochlorite solution for fifteen min followed by three time rising with sterile distil-water. Isolated ovaries were then transferred to MS hormone free medium containing 30 and 60 g/L sucrose for embryo development and germination. Collected data have been analyzed by two factorial experimental designs. The results indicated that there were significant differences between age of embryos for development and germination at ${\alpha}=0.01%$. Highest embryo germination (45%) was observed from 21 days old ovaries. No significant differences were observed between MS culture media containing 30 and 60 g/L for percentages of ovary-embryo germination and number of germinated embryo per ovary at ${\alpha}=0.05%$. Fourteen percentage of embryo germination obtained in MS medium supplemented with 60 g/L sucrose, while only 35% of isolated ovaries were able to germinate in MS containing 30 g/L sucrose. Induced plantlets in 4 cm height were transferred into pots containing soilless (1:1:1 peat, per lit and vermiculite) medium for acclimatization. After successful acclimatization, plants were delivered to nursery.

• 생명과학회지
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• 제13권4호
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• pp.522-528
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• 2003

본 연구는 출아형 효모 Saccharomyces cerevisiae에서 자외선의 상해 시 이를 정상으로 회복시키는 절제회복 (excision repair) 유전자로 알려진 RADA4의 특성 규명을 위하여 균류 Coprinus cinereus에서 이와 유사한 유전자를 분리하였다. RAD4 유사 유전자를 분리하기 위하여 균류 C. cinereus의 염색체 DNA를 전기영동하여 분리한 다음 효모 RAD4 DNA를 probe로하여 이와 hybridization하였다. 이 결과 RAD4 유사 유전자는 3.2 kb의 insert DNA를 갖고 있었다. 또한 Southern hybridization으로 이 유사 유전자는 fungus C. cinereus의 염색체에 존재함을 확인하였다. 분리한 RAD4 유사 유전자의 전사체 크기는 2.5 kb 였으며, 자외선의 상해 시 전혀 'inducibility가 없음을 Northern hybridization으로 확인하였다. 또한 유사유전자 부분을 삭제하였을 때 이 부분이 없는 세포는 전혀 생존을 못하였다. 이 결과 분리한 RAD4 유사유전자는 세포의 생존에 관여함을 알 수 있었다.

• Clinical and Experimental Pediatrics
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• 제46권2호
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• pp.167-172
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• 2003

목 적 : 소아 심장병의 주종을 이루고 있는 선천성 심장병 환아들에서 폐동맥 고혈압은 비교적 흔히 발생하지만 매우 치료하기 어려운 합병증이다. 폐동맥 고혈압의 원인과 치료 및 예방에 대해서는 아직 많이 알려지지 않은 실정이므로 이의 원인을 산소결핍이라는 전형을 이용하여 VEGF란 유전인자의 차원에서 규명하고, 나아가서는 폐동맥 고혈압의 치료 및 예방책을 마련하기 위하여 이 연구를 시행하였다. 방 법 : 폐동맥 평활근 세포는 생후 6주 Fischer rat의 주폐동맥을 적출하여 작은 조각으로 잘라 20% fetal bovine serum을 첨가한 DMEM 배지를 사용하여 5% 이산화탄소 배양기에서 배양하였다. 배양된 세포는 평활근 세포에만 선택적으로 염색되는 평활근 myosin과 ${\alpha}$-actin 항체를 이용하여 염색함으로써 순수 평활근 세포임을 확인하였다. 5% 이산화탄소 배양기에서 배양한 대조군 세포와 1 또는 3% $O_2$ tension에서 배양한 실험군 세포에서의 VEGF 발현 차이와 starvation한 군과 하지 않은 군에서의 VEGF 발현 차이를 RT-PCR과 northern blotting을 이용하여 비교하였다. 결 과 : 대조군과 저산소 조건에서 배양한 실험군에서 VEGF 발현 정도는 차이가 없었다. 결 론 : 아직 국내에서는 유전인자 차원에서의 폐동맥고혈압의 원인규명이나 이에 따른 치료에 대한 연구가 전혀 없는 상태이며, 이 연구에 이어 신생쥐와 성숙쥐와의 차이점 및 나아가서 사람과 쥐의 폐동맥 평활근 세포의 차이점 등을 규명할 예정이며, 이번 연구 결과를 바탕으로 폐동맥 고혈압의 원인기전 규명, 치료 및 예방방법 개발에 기여하고자 한다.

• 한국동물학회지
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• 제39권1호
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• pp.115-121
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• 1996

임신중기에서 발기에 이르는 흰쥐의 태반에서 Placental Lactogen I (PL-I), II 그리고 Pit-1 의 유전자 발현 변화를 Northern blot hybridization으로 조사하였다. 그 결과, 임신시기에 따라 PL-I과 PL-II의 mRNA 양과 크기에 변화가 나타났다. 이들 유전자 발현에 미치는 에스트로겐의 영향을 조사하기 위하여, 임신 14일째 쥐의 난소를 제거하고(OVX), 이후 매일 에스트로겐을 투여한 후 (OVX+E), 임신 18일째 태반을 회수하여 PL-I, II 그리고 Pit-1의 유전자 발현을 Northern blot hybridization으로 조사하였다. OVX 군의 경우, PL-I의 mRNA 크기는 1 kb에서 1.3 kb로 PL-II의 mRNA는 0.6kb에서 1 kb로 변화하였다. OVX+E 군에서는 PL-I과 PL-II의 mRNA가 정상대조군과 같은 상태로 환원하였다. 정상대조군에 비하여 OVX와 OVX+E 군에서 PL-I과 PL-II의 mRNA 크기에는 영향을 미치지 못한 반면, mRNA 양은 난소제거시 감소하였다가, 에스트로겐을 투여하면, 부분적으로 회복되는 경향을 보였다. 이러한 본 실험의결과는 에스트로겐이 PL-I과 PL-II의 RNA splicing이나 polyadenylation 등을 포함한 유전자 발현에 영향을 미치며, Pit-1이 이 과정에 개입하는 것을 시사하는 것이다.

• Fisheries and Aquatic Sciences
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• 제2권2호
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• pp.176-181
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• 1999

The variations of gene expression and pituitary contents of GTH subunits during the ovarian development of masu salmon, Oncorhynchus masou, were investigated. The pituitary GTHs contents was measured by radioimmunoassays (RIAs) using purified GTH subunits and their antibodies. Pituitary contents of GTH $I\beta$ gradually increased from April through August, and reached the maximum in October. On the other hand, pituitary contents of GTH $II\beta$ remained low until August, but they rapidly increased in October. Total RNAs were prepared from pooled pituitaries and the GTH subunits mRNA in pituitaries was quantified by Northern blot hybridization using masu salmon cDNA probes for each GTH subunit. GTH $I\beta$ mRNA level increased with the progression of ovarian maturity. However, GTH $II\beta$ mRNA was detected only at a more advanced stage, and were extremly high at ovulation. A high levels for GTH a mRNA was detected only at ovulation stage. The synchronous increase in pituitary contents and mRNA levels suggested that ovarian maturity in masu salmon was regulated by both GTH I and GTH II.