• KSBB Journal
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• 제21권6호
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• pp.433-438
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• 2006

Bacillus subtilis BB-1 (KFCC l1344P)으로부터 분리된 혈전용해효소 유전자 (BCF-1)를 대량발현 벡터인 pEB 벡터에 크로닝하여 순수분리 된 혈전용해효소를 rat 경구 투여하여 출혈시간, 혈액의 응고, serum의 혈전용해능 등에 대한 in vivo 실험을 실시하였으며, 혈전용해효소의 단회경구투여에 따른 독성을 검사하였다. 효소의 경구투여에 따른 rat의 출혈시간에서는 대조군에 비하여 모든 경구 투여군에서 출혈시간이 유의적으로 약 1.75배 이상 길게 나타남을 확인하였고(P<0.05), 혈액의 출혈시간 또한 활발히 진행됨을 관찰하였다. 혈액으로부터 분리된 serum의 혈전용해작용 있어서는 경구투여 후 1시간부터 채혈한 혈액 내에서 혈전용해효소의 활성이 검출되기 시작하여 3시간째까지 높은 활성을 보였으며 4시간째부터 서서히 활성이 감소하는 것을 확인하였고 혈액의 응고 역시 대조군에 비하여 경구 투여군에서 상당히 지연되는 것을 알 수 있었다. Western blot에 의한 효소 검출에서는 경구 투여군에서 30,000 Da 크기의 단일밴드를 확인하였으며, 혈전용해효소의 rat에 대한 단회경구투여 독성실험에서 중량의 변화, 장기의 이상여부, 사망률 등에서 어떠한 이상이나 병변이 발견되지 않았다. 이상의 결과로 동물실험을 통한 혈전용해 효소의 경구투여에 의한 작용을 혈액 내에서 확인 할 수 있었으며, 본 효소의 단회 경구투여 시의 독성은 전혀 없음을 확인할 수 있었다.

• International Journal of Oral Biology
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• 제37권2호
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• pp.43-50
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• 2012

The use of high throughput screening (HTS) in drug development is principally for the selection new drug candidates or screening of chemical toxicants. This system minimizes the experimental environment and allows for the screening of candidates at the same time. Umbilical cord-derived stem cells have some of the characteristics of fetal stem cell and have several advantages such as the ease with which they can be obtained and lack of ethical issues. To establish a HTS system, optimized conditions that mimic typical cell culture conditions in a minimal space such as 96 well plates are needed for stem cell growth. We have thus established a novel HTS system using human umbilical cord derived-mesenchymal stem cells (hUC-MSCs). To determine the optimal cell number, hUC-MSCs were serially diluted and seeded at 750, 500, 200 and 100 cells per well on 96 well plates. The maintenance efficiencies of these dilutions were compared for 3, 7, 9, and 14 days. The fetal bovine serum (FBS) concentration (20, 10, 5 and 1%) and the cell numbers (750, 500 and 200 cells/well) were compared for 3, 5 and 7 days. In addition, we evaluated the optimal conditions for cell cycle block. These four independent optimization experiments were conducted using an MTT assay. In the results, the optimal conditions for a HTS system using hUC-MSCs were determined to be 300 cell/well cultured for 8 days with 1 or 5% FBS. In addition, we demonstrated that the optimal conditions for a cell cycle block in this culture system are 48 hours in the absence of FBS. In addition, we selected four types of novel small molecule candidates using our HTS system which demonstrates the feasibility if using hUC-MSCs for this type of screen. Moreover, the four candidate compounds can be tested for stem cell research application.

• BMB Reports
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• 제43권6호
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• pp.432-437
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• 2010

LBH is a transcription factor as a candidate gene for CHD associated with partial trisomy 2p syndrome. To identify potential LBH-interacting partners, a yeast two-hybrid screen using LBH as a bait was performed with a human heart cDNA library. One of the clones identified encodes ${\alpha}B$-crystallin. Co-immunoprecipitation and GST pull-down assays showed that LBH interacts with ${\alpha}B$-crystallin, which is further confirmed by mammalian two-hybrid assays. Co-localization analysis showed that in COS-7 cells, ${\alpha}B$-crystallin that is cytoplasmic alone, accumulates partialy in the nucleus when co-transfected with LBH. Transient transfection assays indicated that overexpression of LBH or ${\alpha}B$-crystallin reduced the transcriptional activities of p53 and p21, respectively, Overexpression of both ${\alpha}B$-crystallin and LBH together resulted in a stronger repression of the transcriptional activities of p21 and p53. These results showed that the interaction of LBH and ${\alpha}B$-crystallin may inhibit synergistically the transcriptional regulation of p53 and p21.

• Journal of Plant Biotechnology
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• 제38권4호
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• pp.241-250
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• 2011

식물유의 거의 대부분은 triacylglycerol (TAG) 형태로 종자에 축적되어있으며 이는 종자가 발아할 때에 필수적인 에너지공급원이자 동물과 인간들에게 필수지방산과 중요한 에너지원이다. 최근 식용유의 건강기능성으로 수요증가와 더불어 바이오디젤과 산업원료 등의 산업적 수요도 증가함에 따라 더욱 중요한 자원이 되고 있다. 그래서 생명공학기술로 종자유의 함량을 증진하고자 하면 지질 생합성에 탄소의 유입에 관여하는 조절 유전자를 과발현 또는 억제하는 것이 결정적으로 중요하다. 본 총설에서는 지질함량에 영향을 미치는 것으로 여겨지는 후보 유전자들에 대해 기술하고 이들의 지방 함량 증대 가능성을 조사하였다. 식물의 지방산의 생합성과 종자유의 축적에 관여하는 유전자들은 크게 구분하자면 첫째, TAG가 생합성되기 위해 필요한 전구체를 합성하는 유전자, 둘째, 지방산합성과 TAG 축적에 관여하는 유전자, 셋째, 종자 발달과 종자유 축적에 관여하는 전사인자 유전자가 있다. 종자유 함량을 결정하는 대사들은 앞에서 언급했듯이 매우 복잡하기 때문에 최근에 전사인자의 조절이 다수의 지방생산 대사 유전자를 동시 조작하여 형질전환 식물에서 종자유 함량이 증진하는 것보다 더 바람직한 접근법으로 여겨지고 있다. 그러나 전사조절유전자의 과발현에 의해 나쁜 농업형질의 유도 같은 문제점도 해결해야 한다.

• 생명과학회지
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• 제18권9호
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• pp.1225-1229
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• 2008

본 연구는 한우의 5번 염색체 내에 존재하고 있는 10개의 microsatellite marker의 대립유전자 빈도를 이용하여 한우의 경제형질과의 연관성을 지닌 좌위를 탐색하기 위하여 실시하였다. 한우 326두를 대상으로 10개의 유전좌위를 분석한 결과 총 169개의 대립유전자가 검출되었다. 모든 유전좌위에서 다형성이 검출되었으며 각 유전좌위 별 대립유전자의 수는 9에서 28개로 나타났으며 평균 16.9로 나타났다. 가장 높은 PIC 값을 가진 유전좌위는 DIK2400(0.908)이었으며 가장 낮은 유전좌위는 DIK2718 (0.603)으로 검출되었다. 관측된 이형접합도에서는 DIK1048 (0.655)가 가장 높게 나타났으며, DIK2400 (0.906)은 가장 낮은 것으로 검출되었다. 분석된 MS marker들의 대립유전자의 빈도를 이용하여 경제형질과의 연관성을 탐색하기 위하여 각 경제형질별 육종가를 대상으로 상위그룹과 하위그룹으로 나누어 Chi-square검정을 실시하였다. 분석결과 DIK2828의 239 대립유전자는 근내지방도에서 상위그룹과 하위그룹의 빈도차가 유의적인 차이를 보이는 것으로 나타났다. BMC1009의 279 대립유전자는 도체중과 등지방두께에서 유의적인 차이를 확인하였으며 285대립유전자는 도체중, 등지방두께 그리고 근내지방도에서 유의적인 차이가 확인되었다. DIK4329의 200대립유전자는 등심단면적과 등지방두께에서 유의적인 차이가 확인되었다. 본 연구에서 유의적인 차이가 확인된 유전자좌위는 5번 염색체 내 20 (DIK2828), 41 (BMC1009) 그리고 95 (DIK4329) cM로 나타났다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.98-98
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• 2017

Leaf senescence is the process of aging in plants. Chlorophyll degradation during leaf senescence has the important role translocating nutrients from leaves to storage organs. The functional stay-green with slow leaf yellowing and photosynthesis activity maintenance has been considered one of strategy for increasing crop productivity. Here, we have identified two QTLs on chromosome 9 and 10 for leaf senescence with chlorophyll content of RIL population derived from a cross between Hanareum 2, early leaf senescence Indica-type variety, and Unkwang, delayed leaf senescence Japonica variety. Among these QTLs, we chose qPLS1 QTL on chromosome 9 for further study. qPLS1 was found to explain 14.4% of the total phenotypic variation with 11.2 of LOD score. Through fine-mapping approach, qPLS1 QTL locus was narrowed down to about 25kb in the marker interval between In/del-4-7-9 and In/del-5-9-4. There are 3 genes existed within 25kb of qPLS1 locus: LOC_Os09g36200, LOC_Os09g36210, and LOC_Os09g36220. Among these genes, transcript level of LOC_Os09g36200 was increased during the leaf senescence stage and the expression level of LOC_Os09g36200 in Indica was higher than in Japonica. Finally, we chose LOC_Os09g36200 as candidate gene and renamed it as OsPLS1-In and OsPLS1-Jp from Indica- and Japonica-type rice, respectively. OsPLS1-In and OsPLS1-Jp overexpressing transgenic plants showed both early leaf senescence phenotype. These results indicate that OsPLS1 functions in chlorophyll degradation and the difference of expression level of OsPLS1 cause the difference of leaf senescence between Indica and Japonica in rice.

• Asian-Australasian Journal of Animal Sciences
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• 제29권11호
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• pp.1555-1561
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• 2016

Shank skin color of Korean native chicken (KNC) shows large color variations. It varies from white, yellow, green, bluish or grey to black, whilst in the majority of European breeds the shanks are typically yellow-colored. Three shank skin color-related traits (i.e., lightness [$L^*$], redness [$a^*$], and yellowness [$b^*$]) were measured by a spectrophotometer in 585 progeny from 68 nuclear families in the KNC resource population. We performed genome scan linkage analysis to identify loci that affect quantitatively measured shank skin color traits in KNC. All these birds were genotyped with 167 DNA markers located throughout the 26 autosomes. The SOLAR program was used to conduct multipoint variance-component quantitative trait locus (QTL) analyses. We detected a major QTL that affects $b^*$ value (logarithm of odds [LOD] = 47.5, $p=1.60{\times}10^{-49}$) on GGA24 (GGA for Gallus gallus). At the same location, we also detected a QTL that influences $a^*$ value (LOD = 14.2, $p=6.14{\times}10^{-16}$). Additionally, beta-carotene dioxygenase 2 (BCDO2), the obvious positional candidate gene under the linkage peaks on GGA24, was investigated by the two association tests: i.e., measured genotype association (MGA) and quantitative transmission disequilibrium test (QTDT). Significant associations were detected between BCDO2 g.9367 A>C and $a^*$ ($P_{MGA}=1.69{\times}10^{-28}$; $P_{QTDT}=2.40{\times}10^{-25}$). The strongest associations were between BCDO2 g.9367 A>C and $b^*$ ($P_{MGA}=3.56{\times}10^{-66}$; $P_{QTDT}=1.68{\times}10^{-65}$). However, linkage analyses conditional on the single nucleotide polymorphism indicated that other functional variants should exist. Taken together, we demonstrate for the first time the linkage and association between the BCDO2 locus on GGA24 and quantitatively measured shank skin color traits in KNC.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8893-8900
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• 2014

Dysregulated expression of microRNAs (miRNAs) has been shown to be closely associated with tumor development, progression, and carcinogenesis. However, their clinical implications for gastric cancer remain elusive. To investigate the hypothesis that genome-wide alternations of miRNAs differentiate gastric cancer tissues from those matched adjacent non-tumor tissues (ANTTs), miRNA arrays were employed to examine miRNA expression profiles for the 5-pair discovery stage, and the quantitative real-time polymerase chain reaction (qRTPCR) was applied to validate candidate miRNAs for 48-pair validation stage. Furthermore, the relationship between altered miRNA and clinicopathological features and prognosis of gastric cancer was explored. Among a total of 1,146 miRNAs analyzed, 16 miRNAs were found to be significantly different expressed in tissues from gastric cancer compared to ANTTs (p<0.05). qRT-PCR further confirmed the variation in expression of miR-193b and miR-196a in the validation stage. Down-expression of miR-193b was significantly correlated with Lauren type, differentiation, UICC stage, invasion, and metastasis of gastric cancer (p<0.05), while over-expression of miR-196a was significantly associated with poor differentiation (p=0.022). Moreover, binary logistic regression analysis demonstrated that the UICC stage was a significant risk factor for down-expression of miR-193b (adjusted OR=8.69; 95%CI=1.06-56.91; p=0.043). Additionally, Kaplan-Meier survival curves indicated that patients with a high fold-change of down-regulated miR-193b had a significantly shorter survival time (n=19; median survival=29 months) compared to patients with a low fold-change of down-regulated miR-193b (n=29; median survival=54 months) (p=0.001). Overall survival time of patients with a low fold-change of up-regulated miR-196a (n=27; median survival=52 months) was significantly longer than that of patients with a high fold-change of up-regulated miR-196a (n=21; median survival=46 months) (p=0.003). Hence, miR-193b and miR-196a may be applied as novel and promising prognostic markers in gastric cancer.

• Asian Pacific Journal of Cancer Prevention
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• 제17권sup3호
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• pp.179-183
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• 2016

Breast cancer is the most prevalent type of cancer among women around the world, and mortality is primarily caused by micro-metastatic disease. The complex mechanisms of breast cancer invasion and metastasis are intrinsically related to the malignant cell type so that early detection of micro-metastases can help prolongation of survival for patient. The aim of the present research work was evaluation of the expression status of mammoglobin protein as a candidate molecular marker in the negative sentinel lymph node (SLN). Fifty tumor specimens, and 50 normal adjacent breast tissue samples from the same patients were selected on the basis of having more than 10% tumor content for RNA extraction from SLNs. Tumor samples and normal adjacent breast tissue were archived in the form of frozen fresh tissue in liquid nitrogen. Real-time PCR was performed on a Bioner life express gradient thermal cycler system. Mammoglobin gene overexpression in breast cancer metastasis was investigated. Single marker results were mammaglobin 66.7% and CK19 50.0%, with 58.3% for the two in combination. Due to improved outcome with at least 3 genes (83.3%), it seems, triple marker evaluation will be most likely useful for detecting micro-metastases instead of studying separate genes.

• Journal of Yeungnam Medical Science
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• 제24권2호
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• pp.252-261
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• 2007

비만은 서구와 우리나라에서 가장 흔한 영양 관련 질환이며, 유전적, 환경적, 생활습관인자들에 의해 초래된다. 이 연구에서는 성인750명을 대상으로 ${\beta}_3AR$ Trp64Arg, UCP2 -866G/A 유전자 다형성을 PCR-RFLP법으로 조사한 후, 체질량지수와 비교하여 어떤 유전자의 다형성이 비만과 관련 있는지 분석하였다. ${\beta}_3AR$의 Trp64Arg의 다형성 분포는 Arg64Arg의 경우 체질량지수 27이상인 비만인에서만 관찰되었지만, 전체적으로 비만인과 정상인에서 통계적으로 유의한 차이가 없었다. UCP2 -866G/A 다형성의 분포는 G/A의 경우 비만인 에서, A/A의 경우 정상인에서 더 통계적으로 유의하게 높았다(p<0.05). 이상의 결과로부터 ${\beta}_3AR$ Trp64Arg 다형성에 비해 UCP2 -866G/A 다형성이 한국인에서 비만과 비만관련 질병을 예측하는데 더 유용한 것으로 판단된다.