• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2019년도 추계학술대회
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• pp.98-98
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• 2019

제니(薺苨, Adenophorae Remotiflori Radix)는 "대한민국약전외한약(생약)규격집(KHP)"에 모시대(Adenophora remotiflorus Miquel)의 뿌리로 수재되어있으나, 형태학적으로 유사한 잔대(A. triphylla), 당잔대(A. stricta) 및 더덕(Codonopsis lanceolata)과 오 혼용 우려가 있어 이들을 구별하기 위한 정확하고 객관적인 종 감별법이 필요하다. 본 연구에서는 '제니'의 기원인 모시대와 오 혼용 우려가 있는 종들을 구별 할 수 있는 유전자 마커를 개발하기 위하여 Genbank에 등록된 ycf2 구간을 활요하여 모시대와 잔대, 당잔대를 구분 할 수 있는 INDEL (insertion/deletion) 마커를 개발하였다. 또한, 보다 정확한 종감별을 위해 DNA 바코드로 활용되고 있는 유전자 부위의 염기서열을 분석하여 ITS (25%), atpB-rbcL (15%), atpF-atpH (14%), rpl16 (13%), trnL-F (10%), matK (9%), rpoC1 (7%)에서 변이율(percent of variable sites)을 확인하였다. 향후, 본 연구에서 개발된 INDEL 마커와 더불어 추가적으로 개발을 진행 중인 분자 마커는 한약재 '제니'의 품질관리에 활용 가능할 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권7호
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• pp.3093-3097
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• 2014

Background: Cancer is a major threat to the public health whether in developed or in developing countries. As the most common primary malignant tumor, the morbidity and mortality rate of lung cancer continues to rise in recent ten years worldwide. Chemotherapy is one of the main methods in the treatment of lung cancer, but this is hampered by chemotherapy drug resistance, especially MDR. As a component of the 60S large ribosomal subunit, ribosomal protein L39-L gene was reported to be expressed specifically in the human testis and human cancer samples of various tissue origins. Materials and Methods: Total RNA of cultured drug-resistant and susceptible A549 cells was isolated, and real time quantitative RT-PCR were used to indicate the transcribe difference between amycin resistant and susceptible strain of A549 cells. Viability assay were used to show the amycin resistance difference in RPL39-L transfected A549 cell line than control vector and null-transfected A549 cell line. Results: The ribosomal protein L39-L transcription level was 8.2 times higher in drug-resistant human lung cancer A549 cell line than in susceptible A549 cell line by quantitative RT-PCR analysis. The ribosomal protein L39-L transfected cells showed enhanced drug resistance compared to plasmid vector-transfected or null-transfected cells as determined by methyl tritiated thymidine (3H-TdR) incorporation. Conclusions and Implications for Practice: The ribosomal protein L39-L gene may have effects on the drug resistance mechanism of lung cancer A549 cells.

• Journal of Ginseng Research
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• 제44권1호
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• pp.135-144
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• 2020

Background: Panax species are important herbal medicinal plants in the Araliaceae family. Recently, we reported the complete chloroplast genomes and 45S nuclear ribosomal DNA sequences from seven Panax species, two (P. quinquefolius and P. trifolius) from North America and five (P. ginseng, P. notoginseng, P. japonicus, P. vietnamensis, and P. stipuleanatus) from Asia. Methods: We conducted phylogenetic analysis of these chloroplast sequences with 12 other Araliaceae species and comprehensive comparative analysis among the seven Panax whole chloroplast genomes. Results: We identified 1,128 single nucleotide polymorphisms (SNP) in coding gene sequences, distributed among 72 of the 79 protein-coding genes in the chloroplast genomes of the seven Panax species. The other seven genes (including psaJ, psbN, rpl23, psbF, psbL, rps18, and rps7) were identical among the Panax species. We also discovered that 12 large chloroplast genome fragments were transferred into the mitochondrial genome based on sharing of more than 90% sequence similarity. The total size of transferred fragments was 60,331 bp, corresponding to approximately 38.6% of chloroplast genome. We developed 18 SNP markers from the chloroplast genic coding sequence regions that were not similar to regions in the mitochondrial genome. These markers included two or three species-specific markers for each species and can be used to authenticate all the seven Panax species from the others. Conclusion: The comparative analysis of chloroplast genomes from seven Panax species elucidated their genetic diversity and evolutionary relationships, and 18 species-specific markers were able to discriminate among these species, thereby furthering efforts to protect the ginseng industry from economically motivated adulteration.

• Asian-Australasian Journal of Animal Sciences
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• 제27권4호
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• pp.471-478
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• 2014

Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (GAPDH, ACTB, B2M, RPL4, TBP, and PPIA) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked B2M and PPIA as the best combination for gene expression normalization, while according to NormFinder, TBP was ranked as the most stable reference gene, followed by B2M and PPIA. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, TBP, PPIA and B2M, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.

• Genomics & Informatics
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• 제8권3호
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• pp.108-115
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• 2010

Hypertension is the most prevalent disease worldwide and is itself a risk factor for cerebral, cardiac, and renal diseases. The inconsistency of candidate genes suggested by previous genomewide association studies (GWASs) may be due to not only differences in study design and genetic or environmental background but also the difference in the power of analysis between continuous traits and discrete traits. We analyzed 352,228 single nucleotide polymorphisms (SNPs) in 8842 unrelated Koreans obtained from Ansan and Ansung cohorts. We performed a series of GWA analyses using three different phenotype models; young hypertensive cases (278 subjects) versus elderly normotensive controls (680 subjects); the upper 25% (2211 hypertensive cases) versus the lower 25% of the SBP distribution (2211 hypotensive controls); and finally SBP and DBP as continuous traits (8842 subjects). The numbers of young hypertensive cases and elderly normotensive controls were not large enough to achieve genomewide significance. The model comparing the upper 25% subjects to the lower 25% of subjects showed a power that was approximate to that of QTL analysis. Two neighboring SNPs of the ATP2B1 gene, rs17249754 (SBP, p=$2.53^{-10}$; DBP, p=$1.28{\times}10^{-8}$) and rs7136259 (SBP, p=$1.30{\times}10^{-9}$; DBP, p=$6.41{\times}10^{-8}$), were associated with both SBP and DBP. Interestingly, a SNP of the RPL6 gene, rs11066280, revealed a significant genomewide association with SBP in men only (p=$3.85{\times}10^{-8}$), and four SNPs located near the MAN2A1 gene showed a strong association with DBP only in elderly men aged 60-70 years (e.g., rs6421827, p=$4.86{\times}10^{-8}$). However, we did not observe any gene variant attaining genomewide significance consistently in the three phenotype models except for the ATP2B1 gene variants. In general, the association signal with blood pressure was stronger in women than in men. Genes identified in GWASs are expected to open the way for prevention, early diagnosis, and personalized treatment of hypertension.

• 한국콘텐츠학회논문지
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• 제10권5호
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• pp.343-350
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• 2010

체외충격파쇄석술(Extracorporeal shock wave lithotripsy ; ESWL) 시 환자가 받는 방사선 피폭선량을 측정하기 위하여 신장 및 요관 결석으로 진단을 받은 총 55명(남:36명, 여:19명)을 대상으로 방사선피폭선량을 측정하였다. 측정 방법은 투시 관전압 80kVp, 관전류 5mA로 고정하여 인체 모형의 Rando Phantom과 형광유리 선량계를 사용하였으며, 주요 장기인 양측 신장, 방광, 간에 5분과 10분씩 각각 2회 흡수선량을 측정하여 유효선량으로 환산하였다. 환자 당 평균 시행 횟수는 1.8회(1~4)이었고, 평균 투시시간은 533초(248~2516)로 나타났다. 우측 신장결석 치료 시 우측 신장의 평균값은 2.458mSv, 좌측 신장은 0.152mSv, 간은 1.404mSv, 방광은 0.019mSv로 측정 되었고, 좌측 신장결석 치료 시 좌측 신장의 평균값은 2.496mSv, 우측 신장은 0.252mSv, 간은 0.178mSv, 방광은 0.017mSv이었으며, 하부요관 결석치료 시 방광에서의 평균값은 3.742mSv, 우측 신장은 0.009mSv, 좌측 신장은 0.01mSv로 유효선량이 측정 되었다.

• 생약학회지
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• 제53권4호
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• pp.226-233
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• 2022

Ephedra herbs are defined as stem of Ephedra sinica , Ephedra intermedia and Ephedra equisetina in the Korean Pharmacopoeia. It is important to use pure herbs to derive the safety and efficacy of herbal medicine. However, the identification of these herbs by conventional taxonomic methods is difficult. Recently, many studies have applied these DNA barcoding for the identification of herbal medicinal species using standard DNA markers. In this study, we report a case study in which the identification of Ephedra species was done by DNA barcoding. For identification of Ephedra species, 17 samples were collected, and a reference DNA barcode library was developed using 6 markers (rbcL, matK, ITS2, ycf1, ycf3, and rpoC2). To develop KASP-SNP markers, we selected 4 markers (ycf1, ycf3, rpl2, and rbcL), which were able to distinguish three Ephedra species. In the result, the specific markers for each of the three Ephedra were clustered into FAM-positive section, whereas non-targeted plants were clustered either HEX-positive or negative section. Therefore, we have developed KASP assay that allow rapid and easy Ephedra species identification using three KASP markers.

• 한국식품과학회지
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• 제28권4호
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• pp.616-623
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• 1996

음양곽 성분 중에서 icariin, magnoflorine을 분리하고 이를 표준품으로 음양곽 추출액의 이들 성분에 대하여 HPLC를 이용한 정량법을 수립하였으며 이 성분들의 추출효율을 여러 가지 추출조건하에서 검토하고 그 함량을 분석 하였다. Icariin은 0.1-0.4 mg/ml 농도에서, magnoflorine은 0.02-0.1 mg/ml 농도에서 아래와 같은 조건으로 정량 분석하였다. Icariin의 HPLC조건 :칼럼, LiChrosorbRPl8; 용출용매, MeOH/water/acetic acid (60 : 40 : 0.5); 검출기, 자외부 흡광광도계; 측정파장, 270 nm; 주사량, $25\;{\mu}l$. Icariin의 retention time은 7.37 min으로 다른 성분과도 잘 분리되었다. Icariin 0.1-0.4 mg/ml 50% (v/v)에탄올 용액에 대해 검량선을 작성하여(n=8) 직선을 얻었으며 그 회귀선은 y = -1.4+18296x이고 correlation coefficient (r)=0.9999로 1.00에 매우 접근하였다. Magnoflorine의 HPLC 조건 : 칼럼, LiChrosorb RP18; 용출용매, MeOH/80 mM $Na_{2}\;HPO_{4}/water$ (35 : 35 : 30); 검출기, 자외부 흡광광도계; 측정파장, 270nm ; 주사량, $25\;{\mu}l$. Magnoflorine의 retention time은 11 min이고 $2{\sim}10\;mg/100\;ml$ 50% 에탄올용액을 표준액으로 검량선을 작성하여 직선상의 검량선을 얻었으며(n=6) 그 회귀직선은 y = -0.02+5.7X이고 correlation coefficient (r)=1.0079로 1.00에 근접하였다. 음양곽의 추출용매로 물 또는 50% (v/v)에탄올을 사용하고 추출온도, 추출시간을 달리하였을 때 icariin, magnoflorine, 엑기스의 추출효율을 분석하였다. Icariin은 $50{\%}$ 에탄올($80^{\circ}C$, 1시간)로 추출하였을 때와 물($100^{\circ}C$, 3시간)로 가열 추출하였을 때 거의 같은 정도로 추출되었으나 magnoflorine은 50% 에탄올로 추출하였을 때 더 많이 추출되었다. 반면에 음양곽의 엑기스량은 50% 에탄올로 추출하였을 때 가장 적었고 물로 추출하였을 때는 추출시간이 길면 길수록 더 많았다. 음양곽을 직접 차(茶)로 음용할 때의 추출 조건에서는 icariin 및 magnoflorine의 추출 효율이 약 25%이므로 음양곽 엑기스로 가공한 차제제가 더 바람직하다고 생각된다. 음양곽 중에 icariin및 magnoflorine의 함량은 각각 0.94, 0.16%임을 처음으로 분석하였다.