• Asian-Australasian Journal of Animal Sciences
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• 제27권12호
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• pp.1695-1704
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• 2014

Maternal malnutrition during pregnancy may give rise to female offspring with disrupted ovary functions in adult age. Neonatal ovary development predisposes adult ovary function, yet the effect of maternal nutrition on the neonatal ovary has not been described. Therefore, here we show the impact of maternal protein restriction on the expression of folliculogenic and steroidogenic genes, their regulatory microRNAs and promoter DNA methylation in the ovary of neonatal piglets. Sows were fed either standard-protein (SP, 15% crude protein) or low-protein (LP, 7.5% crude protein) diets throughout gestation. Female piglets born to LP sows showed significantly decreased ovary weight relative to body weight (p<0.05) at birth, which was accompanied with an increased serum estradiol level (p<0.05). The LP piglets demonstrated higher ratio of bcl-2 associated X protein/B cell lymphoma/leukemia-2 mRNA (p<0.01), which was associated with up-regulated mRNA expression of bone morphogenic protein 4 (BMP4) (p<0.05) and proliferating cell nuclear antigen (PCNA) (p<0.05). The steroidogenic gene, cytochrome P450 aromatase (CYP19A1) was significantly down-regulated (p<0.05) in LP piglets. The alterations in ovarian gene expression were associated with a significant down-regulation of follicle-stimulating hormone receptor mRNA expression (p<0.05) in LP piglets. Moreover, three microRNAs, including miR-423-5p targeting both CYP19A1 and PCNA, miR-378 targeting CYP19A1 and miR-210 targeting BMP4, were significantly down-regulated (p<0.05) in the ovary of LP piglets. These results suggest that microRNAs are involved in mediating the effect of maternal protein restriction on ovarian function through regulating the expression of folliculogenic and steroidogenic genes in newborn piglets.

• BMB Reports
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• 제47권5호
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• pp.274-279
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• 2014

Bortezomib has been known as the most promising anti-cancer drug for multiple myeloma (MM). However, recent studies reported that not all MM patients respond to bortezomib. To overcome such a stumbling-block, studies are needed to clarify the mechanisms of bortezomib resistance. In this study, we established a bortezomib-resistant cell line (U266/velR), and explored its biological characteristics. The U266/velR showed reduced sensitivity to bortezomib, and also showed cross-resistance to the chemically unrelated drug thalidomide. U266/velR cells had a higher proportion of CD138 negative subpopulation, known as stem-like feature, compared to parental U266 cells. U266/velR showed relatively less inhibitory effect of prosurvival NF-${\kappa}B$ signaling by bortezomib. Further analysis of RNA microarray identified genes related to ubiquitination that were differentially regulated in U266/velR. Moreover, the expression level of CD52 in U266 cells was associated with bortezomib response. Our findings provide the basis for developing therapeutic strategies in bortezomib-resistant relapsed and refractory MM patients.

• Asian Pacific Journal of Cancer Prevention
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• 제15권4호
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• pp.1817-1822
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• 2014

Objective: The aim of this study was to screen for possible biomarkers of metastatic osteosarcoma (OS) using a DNA microarray. Methods: We downloaded the gene expression profile GSE49003 from Gene Expression Omnibus database, which included 6 gene chips from metastatic and 6 from non-metastatic OS patients. The R package was used to screen and identify differentially expressed genes (DEGs) between metastatic and non-metastatic OS patients. Then we compared the expression of DEGs in the two groups and sub-grouped into up-regulated and down-regulated, followed by functional enrichment analysis using the DAVID system. Subsequently, we constructed an miRNA-DEG regulatory network with the help of WebGestalt software. Results: A total of 323 DEGs, including 134 up-regulated and 189 down-regulated, were screened out. The up-regulated DEGs were enriched in 14 subcategories and most significantly in cytoskeleton organization, while the down-regulated DEGs were prevalent in 13 subcategories, especially wound healing. In addition, we identified two important miRNAs (miR-202 and miR-9) pivotal for OS metastasis, and their relevant genes, CALD1 and STX1A. Conclusions: MiR-202 and miR-9 are potential key factors affecting the metastasis of OS and CALD1 and STX1A may be possible targets beneficial for the treatment of metastatic OS. However, further experimental studies are needed to confirm our results.

• Asian-Australasian Journal of Animal Sciences
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• 제31권10호
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• pp.1550-1557
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• 2018

Objective: Circular RNAs (circRNAs) are a newfound class of non-coding RNA in animals and plants. Recent studies have revealed that circRNAs play important roles in cell proliferation, differentiation, autophagy and apoptosis during development. However, there are few reports about muscle development-related circRNAs in livestock. Methods: RNA sequencing analysis was employed to identify and annotate circRNAs from longissimus dorsi of sheep. Reverse transcription followed by real-time quantitative (q) polymerase chain reaction (PCR) analysis verified the presence of these circRNAs. Targetscan7.0 and miRanda were used to analyse the interaction of circRNA-microRNA (miRNA). To investigate the function of circRNAs, an experiment was conducted to perform enrichment analysis hosting genes of circRNAs using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathways. Results: About 75.5 million sequences were obtained from RNA libraries of sheep skeletal muscle. These sequences were mapped to 729 genes in the sheep reference genome. We identified 886 circRNAs, including numerous circular intronic RNAs and exonic circRNAs. Reverse transcription PCR (RT-PCR) and DNA sequencing analysis confirmed the presence of several circRNAs. Real-Time RT-PCR analysis exhibited resistance of sheep circRNAs to RNase R digestion. We found that many circRNAs interacted with muscle-specific miRNAs involved in growth and development of muscle, especially circ776. The GO and KEGG enrichment analysis showed that hosting genes of circRNAs was involved in muscle cell development and signaling pathway. Conclusion: The study provides comprehensive expression profiles of circRNAs in sheep skeletal muscle. Our study offers a large number of circRNAs to facilitate a better understanding of their roles in muscle growth. Meanwhile, we suggested that circ776 could be analyzed in future study.

• Genes and Genomics
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• 제40권11호
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• pp.1213-1223
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• 2018

Pelagic larval dispersal habits influence the population genetic structure of marine mollusk organisms via gene flow. The genetic information of the clam Gomphina aequilatera (short larval stage, 10 days) which is ecologically and economically important in the China coast is unknown. To determine the influence of planktonic larval duration on the genetic structure of G. aequilatera. Mitochondrial markers, cytochrome oxidase subunit i (COI) and 12S ribosomal RNA (12S rRNA), were used to investigate the population structure of wild G. aequilatera specimens from four China Sea coastal locations (Zhoushan, Nanji Island, Zhangpu and Beihai). Partial COI (685 bp) and 12S rRNA (350 bp) sequences were determined. High level and significant $F_{ST}$ values were obtained among the different localities, based on either COI ($F_{ST}=0.100-0.444$, P<0.05) or 12S rRNA ($F_{ST}=0.193-0.742$, P<0.05), indicating a high degree of genetic differentiation among the populations. The pairwise $N_m$ between Beihai and Zhoushan for COI was 0.626 and the other four pairwise $N_m$ values were >1, indicating extensive gene flow among them. The 12S rRNA showed the same pattern. AMOVA test results for COI and 12S rRNA indicated major genetic variation within the populations: 77.96% within and 22.04% among the populations for COI, 55.73% within and 44.27% among the populations for 12S rRNA. A median-joining network suggested obvious genetic differentiation between the Zhoushan and Beihai populations. This study revealed the extant population genetic structure of G. aequilatera and showed a strong population structure in a species with a short planktonic larval stage.

• 한국자원식물학회지
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• 제19권1호
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• pp.161-168
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• 2006

제주도에 자생하는 부채 선인장인 백년초의 기원 규명을 목적으로 ITS primer를 이용하여 685 bp의 ITS 영역을 분리하였다. ITS 영역의 염기서열을 분석한 결과 18S rRNA의 길이는 54 bp, 26S rRNA는 55 bp, ITS1은 193 bp, ITS2는 220 bp로 구성되어 있었다. 백년초 ITS 영역은 기존에 보고된 Cucurbitoideae 식물들의 ITS 영역에 비하여 ITS2 스페이서 영역의 239-254 bp보다는 다소 짧았다. 그러나 이들 스페이서 영역의 GC 함량은 백년초의 경우 ITS1은 66.8%, ITS2의 경우에는 67.7%로 Cucurbitoideae 식물들에서 보다 높은 GC 함량을 나타내었다. 백년초 선인장의 rDNA 영역에 가장 높은 상동성을 나타낸 것은 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)로 95%의 유사도를 나타내었다. 백년초 rDNA Clustal W 프로그램을 이용하여 유연관계를 조사한 결과 같은 Opuntioideae에 속하는 Pereskiopsis porteri(L78037)와 같은 cluster로 분리되었다.

• 생명과학회지
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• 제17권3호통권83호
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• pp.402-406
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• 2007

우리나라에 분포하는 멧토끼 (Lepus coreanus)의 성판별을 위한 분자 표지자를 개발하기 위하여, X, Y 염색체간 상동인 ZFX와 ZFY 유전자들의 성별 이형성에 초점을 맞추어 본 연구를 수행하였다. ZFX와 ZFY 유전자의 인트론 7 영역은 멧토끼의 암수가 구분되는 증폭 양상을 나타내었다. 인트론 7의 길이는 각각 ZFX에서 538, ZFY에서 233-bp로 확인되었다. 특히, ZFX의 인트론 7에서는 RNA-매개성 전위인자 중 한 종이며 토끼의 유전체에서 빈번하게 관찰되는 CSINE2와 유사한 반복서열이 발견되었다. 반면, 반복서열은 ZFY의 인트론 7에서는 관찰되지 않았다. ZFX와 ZFY 유전자의 인트론 7에서 확인된 길이의 차이에 근거하여 중합효소연쇄반응 기법을 이용한 유전자 성판별을 수행하였다. 시험에 이용된 모든 DNA시료들은 ZFX에서 증폭된 공통의 밴드를 가지고 있었다. 이에 반해, 멧토끼 수컷 DNA들은 각각 ZFX와 ZFY에서 증폭된 두 개의 구분되는 밴드들을 나타내었다. ZFX-ZFY 유전자·성판별 결과는 표현형 성별 정보뿐만 아니라 수컷-특이적인 SRY 유전자의 증폭양상과도 일치한 결과와도 정확히 일치하였다. 이상의 결과들은 멧토끼에서 ZFX와 ZFY의 인트론 7 영역간의 성별 이형성은 유전자 성판별을 위한 유용한 유전자 표지자가 될 것으로 사료된다.

• Clinical and Experimental Pediatrics
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• 제53권2호
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• pp.178-183
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• 2010

목 적 : 최근에 macrolide계 항균제에 내성인 M. pneumoniae 균주가 증가한다는 외국의 보고가 있었으며, 국내에서 수행된 한 연구에서도 M. pneumoniae의 macrolide 내성률을 49% 정도로 보고한 바 있다. 이에, 본 연구는 M. pneumoniae 폐렴으로 진단된 소아의 비인두 흡인물에서 M. pneumoniae의 macrolide 계항균제 내성에 연관된 것으로 알려진 유전자 변이 유무를 확인하고, M. pneumoniae의 macrolide계 항균제에 대한 최소억제농도를 측정하기 위한 기초 연구로 M. pneumoniae 배양법을 구축하고자 시행되었다. 방 법 : 2000년과 2003년 M. pneumoniae 감염의 유행기에 급성 호흡기 증상을 주소로 서울대학교 어린이병원과 분당서울대학교병원에서 치료받은 소아 중 혈청학적 검사와 M. pneumoniae PCR을 통해 M. pneumoniae 폐렴으로 진단받은 환아 62명으로 부터 채취하여 $-80^{\circ}C$에 보관되었던 비인두 흡인물을 대상으로 하였다. M. pneumoniae의 23S rRNA domain V의 peptidyl transferase 부위와 ribosomal protein L4를 M. pneumoniae 특이 PCR로 증폭한 후 염기서열분석을 시행하였다. 염기서열의 분석은 M. pneumoniae 표준 균주와 비교하여, 23S rRNA domain V의 A2063G, A2064G 변이와 ribosomal protein L4의 M144V변이 유무를 확인하였다. 또한, M. pneumoniae 표준 균주와 33개의 비인두흡인물($-80^{\circ}C$에 보관되었던 28검체와 1-2일간 냉장보관되었던 비인두흡인물 5 검체)을 Chanock's glucose 액체배지와 한천배지에 접종하고 $37^{\circ}C$의 5% $CO_2$ 항온기에서 6주간 관찰하여 배양을 확인하였다. 결 과 : 총 62 검체 중 23S rRNA gene에 대한 염기서열분석이 가능했던 61 검체 중 1검체(1.6%)에서 A2064G변이가 관찰되었고, 62 검체의 ribosomal protein L4에 대한 염기서열분석 결과 17검체(27.4%)에서 M144V 아미노산 변이가 확인되었다. M. pneumoniae 배양 결과, 표준 균주는 Chanock's glucose 액체배지와 한천배지 모두에서 배양되었고 2009년에 채취된 5검체 중 2검체에서 배양이 확인되었으나, $-80^{\circ}C$에 보관되었던 28검체는 모두 배양되지 않았다. 결 론 : 본 연구에서 23S rRNA gene의 유전자 변이 빈도는 매우 낮았고, ribosomal protein L4의 M144V 변이는 좀 더 많은 검체에서 확인되었다. Macrolide계 항균제에 내성인 M. pneumoniae의 분포와 M. pneumoniae의 23S rRNA gene과 ribosomal protein L4의 변이에 대한 추가적인 연구들을 통해 M. pneumoniae의 macrolide 항균제에 대한 내성기전을 이해하는데 도움을 줄 수 있을 것으로 생각된다.

• 생명과학회지
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• 제30권1호
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• pp.88-95
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• 2020

유기용매 내성 세균인 Pseudomonas sp. BCNU 106을 10%(v/v) 톨루엔에 노출시킨 후 8시간 동안 random arbitrarily primed polymerase chain reaction (RAP-PCR)기법을 이용하여 메신져 RNA 발현 레벨을 조사하였다. 총 100개의 상향발현된 발현 산물 중에서 50개의 상보적인 단편들이 반복적으로 재현성있게 발현되는 것으로 확인되어, 이들을 클로닝을 하였으며 나아가 유전자 염기서열을 결정하였다. Blast analysis 결과, 톨루엔은 LysR family transcriptional regulator 그리고 RNA polymerase factor sigma-32같은 전사와 관련된 유전자들의 발현 레벨을 적응적으로 증가시키는 것으로 확인되었다. 그리고 톨루엔 스트레스 존재 하에서 inorganic ion 수송과 대사와 관련된 catalase와 Mn²⁺/Fe²⁺ transporter 유전자의 발현이 증가되었으며, 신호전달과 대사와 기능적으로 관련된 type IV pilus assembly PilZ와 multi-sensor signal transduction histidine kinase 유전자들의 발현 증가도 확인되었다. 한편 톨루엔 노출 후 탄수화물 수송과 대사와 관련된 beta-hexosaminidase 유전자발현 레벨이 증가하였다. 나아가 DNA polymerase III subunit epsilon, DNA-3-methyladenine glycosylase II와 DEAD/DEAH box helicase domain-containing 유전자들과 같은 DNA 복제, 재조합 그리고 수복에 관련성이 있는 유전자들의 발현 레벨 그리고 심지어는 ABC transporter 유전자와 같은 방어 메커니즘에 관련성이 있는 유전자들의 발현 레벨이 적응적으로 증가되는 것으로 밝혀졌다. 특히 10% 톨루엔 존재하에서 ABC transportor, Mn²⁺/Fe²⁺ transporter 및 β-hexosaminidase 유전자에 해당하는 RNA들이 괄목하게 유도되는 것이 확인되었다. 그러므로 유기용매 내성 세균 Pseudomonas sp. BCNU 106이 유기용매에 대하여 내성을 나타내는데 있어서 방어 메커니즘, 세포내 이온 항상성 그리고 바이오 필름 형성이 필수적인 것으로 확인되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.576-583
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• 2009

G protein-coupled receptor 43 (GPR43) is a newly-discovered short-chain free fatty acid receptor and its functions remain to be defined. The objective of this study was to investigate the function of GPR43 on lipolysis. We successfully cloned the GPR43 gene from the pig (EU122439), and measured the level of GPR43 mRNA in different tissues and primary pig adipocytes. The expression level of GPR43 mRNA was higher in adipose tissue and increased gradually with adipocyte differentiation. Then we examined GPR43 mRNA level in different types, growth-stages and various regions of adipose tissue of pigs. The results showed that the expression level of GPR43 mRNA was significantly higher in adipose tissue of obese pigs than in lean pigs, and the expression level also gradually increased as age increased. We further found that the abundance of GPR43 mRNA level increased more in subcutaneous fat than visceral fat. Thereafter, we studied the correlation between GPR43 and lipid metabolism-related genes in adipose tissue and primary pig adipocytes. GPR43 gene had significant negative correlation with hormone-sensitive lipase gene (HSL, r = -0.881, p<0.01) and triacylglycerol hydrolase gene (TGH, r = -0.848, p<0.01) in adipose tissue, and had positive correlation with peroxisome proliferator-activated receptor $\gamma$ gene ($PPAR_{\gamma}$, r = 0.809, p<0.01) and lipoprotein lipase gene (LPL, r = 0.847, p<0.01) in adipocytes. In addition, we fed different concentrations of docosahexaenoic acid (DHA) to mice, and analyzed expression level changes of GPR43, HSL and TGH in adipose. The results showed that DHA down-regulated GPR43 and up-regulated HSL and TGH mRNA levels; GPR43 also had significant negative correlation with HSL (low: r = -0.762, p<0.01; high: r = -0.838, p<0.01) and TGH (low: r = -0.736, p<0.01; high: r = -0.586, p<0.01). Our results suggested that GPR43 is a potential factor which regulates lipolysis in adipose tissue, and DHA as a receptor of GPR43 might promote lipolysis through down-regulating the expression of GPR43 mRNA.