• Asian-Australasian Journal of Animal Sciences
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• 제22권6호
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• pp.871-884
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• 2009

Gene expression profiling is a useful tool for identifying critical genes and pathways in metabolism. The objective of this study was to determine the major differences in the expression of genes associated with metabolism and metabolic regulation in liver and mammary tissues of lactating cows. We used the Michigan State University bovine metabolism (BMET) microarray; previously, we have designed a bovine metabolism-focused microarray containing known genes of metabolic interest using publicly available genomic internet database resources. This is a high-density array of 70mer oligonucleotides representing 2,349 bovine genes. The expression of 922 genes was different at p<0.05, and 398 genes (17%) were differentially expressed by two-fold or more with 222 higher in liver and 176 higher in mammary tissue. Gene ontology categories with a high percentage of genes more highly expressed in liver than mammary tissues included carbohydrate metabolism (glycolysis, glucoenogenesis, propanoate metabolism, butanoate metabolism, electron carrier and donor activity), lipid metabolism (fatty acid oxidation, chylomicron/lipid transport, bile acid metabolism, cholesterol metabolism, steroid metabolism, ketone body formation), and amino acid/nitrogen metabolism (amino acid biosynthetic process, amino acid catabolic process, urea cycle, and glutathione metabolic process). Categories with more genes highly expressed in mammary than liver tissue included amino acid and sugar transporters and MAPK, Wnt, and JAK-STAT signaling pathways. Real-time PCR analysis showed consistent results with those of microarray analysis for all 12 genes tested. In conclusion, microarray analyses clearly identified differential gene expression profiles between hepatic and mammary tissues that are consistent with the differences in metabolism of these two tissues. This study enables understanding of the molecular basis of metabolic adaptation of the liver and mammary gland during lactation in bovine species.

• BMB Reports
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• 제42권6호
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• pp.373-379
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• 2009

5-Hydroxyuridine (5-OHU) is a major lesion of uridine and cytosine produced in RNA by various chemical oxidants. To elucidate its biochemical and biophysical effects on RNA replication, the site-specifically modified oligoribonucleotides containing 5-OHU were synthesized with C5-hydroxy-5'-ODMTr-2'-TBDMS-uridine phosphoramidite using automated solid phase synthesis. The base-pairing properties of nucleotides opposite 5-OHU in 24 mer oligoribonulcleotides with dNTP were studied using three reverse transcriptases (Super-$Script^{TM}II$-, AMV-, MMLV-RT) in cDNA synthesis. Adenine as well as guanine was incorporated preferentially by all reverse transcriptases. In the UV-melting temperature experiment, the results from the relative stabilities of the base pairs were A : 5-OHU > G : 5-OHU > T : 5-OHU $\approx$ C : 5-OHU. Circular Dichroism (CD) studies showed that DNA-RNA containing 5- OHU heteroduplexes exhibit a similar conformation between the A-type RNA and B-type DNA. These results suggest that 5- OHU from oxidative damage was mainly influenced by adenine mismatch.

• Journal of Animal Science and Technology
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• 제46권6호
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• pp.897-902
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• 2004

이전에 연구된 한우육과 Holstein 및 Black angus육 감별법의 신속성, 편리성, 경제성 등의 단점을 보완하기 위해 소의 MC1R gene의 SNP를 이용한 새로운 감별법을 시도하였다. 본 연구에서는 소의 MC1R gene 중 594번째 염기인 Guanine이 한우에서는 결실된 점을 이용하여 한우의 sequence를 바탕으로 3 쪽에 2mers의 tail을 달아 한우에게는 상보적이나 다른 종에서는 상보적이지 않은 3 -tailed primers를 제작하였다. 이 primer들을 이용해서 한우에서만 MC1R 중 571번째 염기서열부터 919번째 염기서열까지의 343bp의 단편이 증폭되도록 하였다. 그 결과, Holstein, Black angus에서는 모두 band가 관찰되지 않았으나 한우에서는 343bp의 band가 확인되었다. 따라서 본 연구에서 사용한 3 -tailed primer를 이용하면 보다 정확하고 재현성 있으며 신속하고 편리하며 경제적인 한우육의 감별이 될 것으로 판단된다.

• KSBB Journal
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• 제20권2호
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• pp.88-92
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• 2005

인산화 과정은 serine, threonine, tyrosine에서 발생하는 생화학적 반응으로, 본 연구에서는 다양한 응용의 잠재능력이 있는 phosphoserine, phosphothreonine, phosphotyrosine에 대한 엡타머를 개발하였다. 우선 in vitro selection 방법에 의해 combinatorial chemistry로부터 얻어진 RNA library로부터 이들 phosphoamino acids와 친화도를 가지고 있는 엡타머를 찾아낼 수 있었다. 총 10번의 일련 과정을 통해 phosphoserine에 대해서 2.6 nM의 친화도를 가지고 있는 엡타머를 (SeA-06), phosphothreonine에 대해서는 2.7nM의 친화도를 가지고 있는 엡타머 (TrA-18)를 찾아낼 수 있었고, 이들의 RNA 2차 구조를 각각 예측하여 보았다. 그러나 phosphotyrosine의 경우 짧은 길이의 엡타머가 selection됨으로 내부적으로 구조를 가지는 엡타머는 얻을 수 없었다. o-phosphoserine에 대한 항체가 기존에 보고가 되었으나 유사한 구조를 지닌 o-phosphothreonine에도 비슷한 활성을 보여 이들을 구분할 수 있는 리간드를 찾기 힘들었으나, 본 연구에서는 엡타머를 사용한 특이성 조사에서도 서로를 극명하게 구별할 수 있음을 확인할 수 있었다. 이와 같이 발굴된 엡타머를 사용하여 biochip이나 Proteomics 분석 도구의 응용에 큰 기대효과를 제공할 수 있을 것이라 사료된다.

• Archives of Pharmacal Research
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• 제29권10호
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• pp.890-897
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• 2006

In inflammatory responses, induction of cytokines and other immune regulator genes in macrophages by pathogen-associated signal such as lipopolysaccharide (LPS) plays a crucial role. In this study, the gene expression profile changes by LPS treatment in the macrophage/monocyte lineage cell line RAW264.7 was investigated. A 60-mer oligonucleotide microarray of which probes target 32381 mouse genes was used. A reverse transcription-in vitro translation labeling protocol and a chemileuminescence detection system were employed. The mRNA expression levels in RAW264.7 cells treated for 6 h with LPS and the control vehicle were compared. 747 genes were up-regulated and 523 genes were down-regulated by more than 2 folds. 320 genes showing more than 4-fold change by LPS treatment were further classified for the biological process, molecular function, and signaling pathway. The biological process categories that showed high number of increased genes include the immunity and defense, the nucleic acid metabolism, the protein metabolism and modification, and the signal transduction process. The chemokine-cytokine signaling, interleukin signaling, Toll receptor signaling, and apoptosis signaling pathways involved high number of genes differentially expressed in response to LPS. These expression profile data provide more comprehensive information on LPS-target genes in RAW264.7 cells, which will be useful in comparing gene expression changes induced by extracts and compounds from anti-inflammatory medicinal herbs.

• Journal of Plant Biotechnology
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• 제5권2호
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• pp.83-86
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• 2003

A reverse transcription polymerase chain reaction (RT-PCR) protocol was developed using two specific 22-mer primers located in coat protein gene of SPFMV. A 411 bp PCR-product was detected in virus infected plants as well as tissue culture raised sweet potato but not in healthy plants. For optimization of RT-PCR protocol, the optimum crude nucleic acid concentration, annealing temperature, primer concentration and numbers of PCR-cycle for maximum sensitivity and specificity were determined. The optimum condition for RT-PCR was as follows: RT-PCR reaction mixture was one-step mixture, containing 50 pmol of primer, 30 units of reverse transcriptase, 5 units of RNasin, and the crude nucleic acid extracts (200 ng). In RT-PCR, cDNA was synthesized at $42^{\circ}C$ for 45 min before a quick incubation on ice after pre-denaturation at $95^{\circ}C$ for 5 min. The PCR reaction was carried out for 40 cycles at $96^{\circ}C$ for 30 see, $63^{\circ}C$ for 30 sec, $72^{\circ}C$ for 1 min, and finally at $72^{\circ}C$ for 10 min. The viral origin of the amplified product was confirmed by sequencing, with the sequence obtained having $95-98\%$ homology with published sequence data for SPFMV. The benefits of this RT-PCR based detection of SPFMV would be simple, rapid and specific.

• The Plant Pathology Journal
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• 제39권3호
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• pp.255-264
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• 2023

Sweet potato symptomless virus 1 (SPSMV-1) is a single-stranded circular DNA virus, belonging to the genus Mastrevirus (family Geminiviridae) that was first identified on sweet potato plants in South Korea in 2012. Although SPSMV-1 does not induce distinct symptoms in sweet potato plants, its co-infection with different sweet potato viruses is highly prevalent, and thus threatens sweet potato production in South Korea. In this study, the complete genome sequence of a Korean isolate of SPSMV-1 was obtained by Sanger sequencing of polymerase chain reaction (PCR) amplicons from sweet potato plants collected in the field (Suwon). An infectious clone of SPSMV-1 (1.1-mer) was constructed, cloned into the plant expression vector pCAMBIA1303, and agro-inoculated into Nicotiana benthamiana using three Agrobacterium tumefaciens strains (GV3101, LBA4404, and EHA105). Although no visual differences were observed between the mock and infected groups, SPSMV-1 accumulation was detected in the roots, stems, and newly produced leaves through PCR. The A. tumefaciens strain LBA4404 was the most effective at transferring the SPSMV-1 genome to N. benthamiana. We confirmed the viral replication in N. benthamiana samples through strand-specific amplification using virion-sense- and complementary-sense-specific primer sets.

• 광물과 암석
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• 제34권3호
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• pp.169-176
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• 2021

본 연구는 압력을 이용한 제올라이트 내 중금속 양이온 또는 CO₂ 기체 포집 등의 응용연구를 하기 위한 기초 단계로 제올라이트의 압력 및 압력전달 매개체(Pressure-transmitting medium, PTM)에 따른 결정구조의 변화를 알아보기위한 목적으로 실험을 진행하였다. 천연 제올라이트 멜리노이트(Merlinoite, (K,Ca_0.5,Ba_0.5,Na)₁₀ Al₁₀Si₂₂O₆₄× 22H₂O)와 동일한 골격구조를 가지는 합성 물질인 제올라이트-W(K_6.4Al_6.5Si_25.8O₆₄× 15.3H₂O, K-MER)의 압력 하 압력전달 매개체에 따른 선형 압축률 및 체적 탄성률의 변화에 대한 X선 회절연구를 진행하였다. 합성된 시료는 정방정계에 속하는 I4/mmm 공간군으로 확인되었다. 압력전달 매개체 중 제올라이트의 동공 및 채널을 투과할 수 있는 투과 매개체(Penetrating medium)로 물, 이산화탄소를, 비투과 매개체로 실리콘 오일(Silicone-oil)을 각각 사용하였으며, 상압에서 최대 3 GPa까지 약 0.5 GPa 간격으로 가압하였다. 다이아몬드 고압유도장치 및 방사광 X-선원을 이용하여 압력 하 시료의 분말 회절을 측정하였고, 르바일(Le-Bail)법 및 버치-머내한 상태방정식을 이용하여 격자상수 및 체적탄성률의 변화를 관찰하였다. 모든 실험에서 c축의 선형압축률(𝛽^c)은 0.006(1) GPa^-1또는 0.007(1) GPa^-1의 값을 보여 압력 증가 대비 유사한 압축률을 보인 반면, a축의 압축률(𝛽^a)은 실리콘 오일 실험에서 0.013(1) GPa^-1을 보여 물과 이산화탄소 (𝛽^a=0.006(1) GPa^-1) 실험결과에 비해 압축률이 약 두 배정도 큰 것으로 확인할 수 있었다. 체적탄성률(K₀)은 물, 이산화탄소, 실리콘 오일의 실험에서 각각 50(3) GPa, 52(3) GPa, 29(2) GPa로 도출되었다. 압력 증가에 따른 ac면의 orthorhombicity를 측정한 결과 물과 이산화탄소 실험에서는 0.350~0.353의 비교적 일정한 값을 보였으나, 실리콘 오일의 실험에서는 y = -0.005(1)x + 0.351(1)의 함수를 만족시키며 압력이 증가할수록 값이 점차 작아졌다.

• International Journal of Industrial Entomology and Biomaterials
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• 제8권2호
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• pp.161-167
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• 2004

Silkworms sex determination drew high attention from researchers. Sex chromosomes on the silkworm are of ZW type for females and ZZ type for males. Chromosome W plays an important role in sex determination. Although several molecular linkage maps have been constructed for silkworm, very few markers are discovered on the W chromosome. In order to look for molecular markers and to further locate the Fern gene on chromosome W, we used genomic DNA from both female and male larvae of a silkworm strain named 937 as PCR templates for RAPD amplification with 200 arbitrary 10-mer primers. The amplification results showed three female-specific bands, namely ${OPG-07_496}, {OPC-15_1,660} and {OPE-18_1,279}$. Further verification, however, revealed no band from OPG-07 and OPC-15 in either sex in the strain 798, but OPE-18 provided female-specific band in the strains Suluan7 and C108, and absent in both males and strain 798. This indicates that the bands from ${OPG-07_496} and {OPC-15_1,660}$ are probably female-specific in strain 937, and the band from OPE-18 was probably amplified from a common segment shared by most strains. The genomic DNAs from OPG-07 and OPC-15 were cloned and sequenced. Sequence analysis showed that the DNAs from OPG-07 and OPC-15 have high identities with the retrotransposable elements, and DNA from OPC-15 contains a portion of sequence which probably encodes an eukaryotic translation initiation factor 4E binding protein (eIF4EBP).

• Journal of Microbiology and Biotechnology
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• 제10권4호
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• pp.522-528
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• 2000

A total of about 300 fungal isolates from forest havitats were screened for inhibitors of tobacco mosaic virus (TMV) infection using its local lesion host, Nicotiana tabacum cv. Xanthi nc. Ine of the isolates, 14T, showed a strong activity against TMV infection, and was identified as an Apiocrea sp. based on its morphological characterstics. Rice was an optimum culture medium for its fermentation, and two antiviral compounds, KGT 141 and KGT 142, were resolved from the rice culture through column chromatography, TLC, and HPLC. By NMR and FAB-MS, the two compounds were identified as chrysospermins B (KGT 141) and D (KGT 142), both of which are peptaibols with 19-mer amino acids possessing an acetylated N-terminus and a hydroxy-amino acid (tryptophanol) at the C-terminus. Both compounds showed inhibitory activities against TMV infection, but chrysospermin D showed the stronger activity than chrysospermin B. The former of $100{\;}\mu\textrm{g}/ml$ and 54.7% at $10{\;}\mu\textrm{g}/ml$, respectively. Furthermore, the chrysospermins were highly cytotoxic toward cancer cell lines of PC-3 (prostrate) and K562 (leukemia), and inhibited growth of the Gram-positive bacteria tested, especially the plant pathogenic bacterium Corynebacterium lilium. To the best of our knowledge, this is the first report on the inhibition of plant virus infection by antimicrobial peptaibols.