• Title/Summary/Keyword: DNA 혼성화

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Effects of Retinoic Acid on Differentiation and Gene Expression of Pig Preadipocytes (Retinoic Acid가 돼지 지방전구세포의 분화와 유전자 발현에 미치는 영향)

  • Lim, Hee-Kyong;Choi, Kang-Duk;Oyungerel, Baatartsogt;Choi, Young-Suk;Chung, Chung-Soo
    • Journal of Animal Science and Technology
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    • v.50 no.4
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    • pp.475-484
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    • 2008
  • The current study was undertaken to determine the effect of retinoic acid(RA) on differentiation and gene expression of pig preadipocytes. The preadipocytes were isolated from the backfat of the new-born pigs. RA was treated to the cultured cells for 4 days and RNA was extracted from the cells. Isolated RNA went through in situ hybridization using the 14,688-gene cDNA microarray chip. Degree of cell differentiation was determined by measuring glycerol 3-phosphate dehydrogenase activity. RA decreased differentiation of pig preadipocytes by 78%. Fourteen genes were significantly up-regulated by RA, including genes known to be involved in lipid metabolism, particulary sphingomyelin phosphodiesterase, apolipoprotein R precursor, growth factor receptor-bound protein 14, retinoic acid receptor RXR gamma. However, the expression of vascular endothelial growth factor D precursor and growth hormone receptor precursor genes playing a central role in cell growth, was greatly decreased. These results suggest that RA inhibits differentiation of pig preadiocytes by regulation of gene expression of the growth factor or growth hormone receptor.

Cataloguing of Anther Expressed Genes through Differential Slot Blot in Oriental Lily (Lilium Oriental Hybrid 'Acapulco') (아카풀코나리에서 Differential Slot Blot을 이용한 약발현 유전자 목록작성)

  • Suh, Eun-Jung;Yu, Hee Ju;Han, Bong Hee;Lim, Yong Pyo;Jeong, Mi-Jeong;Lee, Seong-Kon;Kim, Dong-Hern;Chang, An-Cheol;Yae, Byeong Woo
    • Horticultural Science & Technology
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    • v.31 no.5
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    • pp.598-606
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    • 2013
  • Anther is the major organ of flower in responsible to reproduction and outward appearance. From anther-specific cDNA library of Lilium Oriental Hybrid 'Acapulco', 2000 expressed sequence tags were selected randomly. Differential slot blot analysis with cDNA probes from the anther and leaf was used to get anther-expressed clone and 570 non-redundant ESTs were obtained and sequenced. Compared to the GenBank database using BLASTX algorithm, 191 clones showed significant similarity but others (66.5%) did not measured to known sequence. Functional categories according to gene ontology (GO) annotation included sequence representing a significant portion of protein in cell and cell part respectively. A transcriptional analysis at 7 different organs and developmental stage was performed using northern blot with thirty ESTs as putative anther specific gene. This report suggest that selection of anther expressed clone using differential slot blot was considered as very effective tool and our current study can provide fundamental information on the lily anther including pollen furthermore.

Changes in Nodule-Specific Proteins during Nodule Development of Canavalia lineata (해녀콩(Canavalia lineata)의 뿌리혹 발달 단계에 따른 뿌리혹 특이 단백질의 변화 양상)

  • 최성화
    • Journal of Plant Biology
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    • v.34 no.2
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    • pp.121-127
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    • 1991
  • Total soluble proteins from three developmental stages of induced root nodules of Canavalia lineata were compared with those of non-nodulated roots by SDS-PAGE and two dimensional (2-D) gel electrophoresis. Thirteen nodule-specific protein (nodulin) bands were identified by the former and 30 nodule specific protein spots were detected by the latter method respectively. Some of the nodulins were detected differentially depending on the nodule's developmental stages. For example, only three leghemoglobin (Lb)-like protein spots appeared at stage I (d<2 mm), but two additional Lb-like protein spots appeared at stage II (d <4-5 mm). pI value and molecular weight of nomomers of Lb-like protein were narrower and greater than those of soybean, ranging from 4.4 to 5.0 and 15.7 kd respectively. Northern blot hybridization of total RNAs from roots and root nodules using soybean Lb cDNA as a probe made it clear that Lb gene was expressed tissue-specifically only in the root nodules.odules.

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Development of Oligonucleotide Chip for Detection of Drug-Resistant Mycobacterium Tuberculosis (약제내성 결핵균의 검출을 위한 Oligonucleotide Chip의 개발)

  • Song, Eunsil;Park, Heekyung;Jang, Hyunjung;Kim, Hyomyung;Chang, Chulhun L.;Kim, Cheolmin
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.41-58
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    • 2003
  • Background : The resurgence of tuberculosis and the widespread emergence of multidrug-resistant M. tuberculosis have emphasized the importance of rapid and accurate diagnostic procedures. Recently, the oligonucleotide chip has proven to be a useful tool in the rapid diagnosis of infectious diseases. The purpose of this study was to rapidly and accurately detect specific mutations in the rpoB, katG and rpsL genes associated with rifampin, isoniazid and streptomycin resistance in M. tuberculosis, respectively, using a single oligonucleotide chip. Method : For detection of drug-resistance, 7 wild-type and 13 mutant-type probes for rifampin, 2 wild-type and 3 mutant-type probes for isoniazid, and 2 wild-type and 2 mutant-type probes for streptomycin were designed and spotted onto glass slides. Fifty-five cultured samples of M. tuberculosis were amplified by PCR, and then underwent hybridization and scanning. Direct sequencing was done to verify the results from the oligonucleotide chip and to analyze the types of mutations. Result : Thirty-five cases out of 40 rifampin-resistant strains(~88%) had mutations in the rpoB gene. One case had a new mutation(D516F, GAC R TTC) and another known mutation together. Twenty cases out of 42 isoniazid-resistant strains(~50%) had mutations in the katG gene, while 7 cases out of 9 streptomycin-resistant strains(~78%) had mutations in the rpsL gene. From these results, the oligonucleotide chip was confirmed to be able to detect the most frequent mutations from the genes associated with rifampin, isoniazid and streptomycin resistance. The results proved that the drug-resistance detection probes were specific. When the results from the oligonucleotide chip and DNA sequencing were compared, the types of mutations were exactly matched. Conclusion : The diagnostic oligonucleotide chip with mutation specific probes for drug resistance is a very reliable and useful tool for the rapid and accurate diagnosis of drug resistance against rifampin, isoniazid and streptomycin in M. tuberculosis infections.