본 연구에서는 DNA marker가 검정된 한우로부터 생산한 체외수정란을 이식하여 육질 및 육량의 유전적 능력이 우수한 한우를 대량생산하여 고품질 한우 쇠고기 생산 시스템을 구축하기 위한 전단계로서 DNA marker 검정 한우로부터 초음파유도 난포란을 채란하여 체외수성 및 수정란의 체외 발달에 미치는 각종 요인들과 배반포기 수정란의 부화율 개선을 위하여 투명대를 laser로 drilling을 실시하여 부화율을 조사하였다. 초음파유래 체외수정란의 분할률

본 연구는 고품질육의 DNA marker가 규떵된 한우로부터 초음파유노 난포란의 연속적 채취를 통하여 능력이 우수한 한우 수정란온 대량생산하는 방법의 확립과 이를 한우농가에 응용하고자 초음파 난자채취기를 이용하여 등지방층두께, 일당증체량, 근내지방도 및배최장근 단면적에 연관된 DNA marker를 보유하고 있는 한우 5두로부터 개체및 난포수, 채취방법, 회수한 난포란의 등급 등을 조사하였다. 한우 5두의 개체별 난포수는 6, 10, 5, 4 및 11회

유전자를 통해 개인의 질병 · 신체적 특징 정보를 분석하는 "유전자 혁명" 시대가 열리고 있다. 최근 유전자 분석 기술의 발전으로 가격과 분석 기간이 급속도로 줄고 있기 때문이다. 인간은 DNA 내 30억 쌍의 염기서열에 유전자를 담고 있는데, 염기서열을 분석하는 컴퓨터 성능이 크게 발전한 덕분이다. 이러한 시대에 맞춰 정부에서도 유전자 검사에 관한 새로운 상품 및 서비스에 맞는 인증, 허가 기준을 위한 산업융합 규제 샌드박스 제도를 실시하게 되었다. 규제 샌드박스는 신산업・ 신기술 분야에서 새로운 제품이나 서비스를 출시할 때 일정 기간 동안 기존 규제를 면제하거나 유예시켜주는 제도이다. 우리나라에서도 이러한 비용 절감이 우리생활에서 쉽게 할 수 있는 유전자검사로 이어졌고 지난 2016년 6월 ‘생명윤리 및 안전에 관한 법률’이 개정되면서 민간 기업이 일반인을 대상으로 한 유전자 분석 서비스, DTC(Direct to Consumer, 소비자 직접 의뢰) 상품시장이 열렸다. 이에 의료 · 제약업체들이 유전자 검사 서비스를 속속 출시, 다양한 유통채널을 통해 판매하고 있는 모습이다. 이에 국내에 제공되고 있는 개인 유전자 DTC 검사 키트 상품을 경험디자인적 관점에서 분석하여 검사 신청에서 실질적인 검체 채취와 반송에 이르기 까지 가장 적합한 유전자검사 키트 상품 제안은 의미가 크다고 하겠다. 사용자의 경험을 기반으로 한 유전자검사 키트 상품 개발은 앞으로 효율적이고 대중적인 개인 유전자 검사보급에 기초 적 자료 활용이 가능할 것이며 새로운 의료 문화형성과 국민보건의료 향상에 도움을 줄 것으로 기대한다.

Cytochrome c oxidase subunit Ⅱ gene(COⅡ gene) is subunit of cytochrome oxidase, which is complex Ⅳ of mitochondria electron transport system. It has been frequently used in molecular phylogenetic studies because the speed of its DNA variation is faster than that of nucleus. It is especially useful in phylogenetic study of molecular biology in insects. In this study, we cloned and sequenced COⅡ gene of mitochondria DNA from Rhabditidae family nematode. Our results showed that this gene is comprised of 696 base pairs(bp). In the analysis of similarity of this gene with other known genes of 14 species of nematodes in Rhabditida order, we identified that this gene has high similarity with that of Caenorhabditis briggsae(86.0%) and C. elegans(85.6%) in Rhabditidae family. On the meanwhile, it has very low similarity with that of Angiostrongylus cantonensis(31.8%) in Angiostrongylidae family and Metastrongylus salmi(31.6%) in Metastrongylidae family. Based on the results of this study, we suggest that this nematode is closely related with that of Caenorhabditis genus in Rhabditidae family.

Background : Chrysanthemi Indici Flos (甘菊) is listed in 「The Korea Herbal Pharmacopoeia (KHP)」as the original plant of Chrysanthemum indicum L. C. indicum was one of the most representative medicinal plants in Asteraceae, Dried flowers of this plant have been valid chemical composition such as flavonoids, phenylpropanoids, terpenoids, and polysaccharides, possessing broad spectrum antibacterial, antiviral, antihypertensive and anti-oxidation functions. Meanwhile, C. indicum was a polymorphic species, its morphological characteristics showed great diversity due to the different geographical and environmental factors. For this reason, there was conducted to develop molecular markers to distinguishing these C. indicum with C. morifolium, C. zawadskii var. latilobum and Aster spathulifolius by using conventional polymerase chain reaction (PCR). Methods and Results : In this study, In order to clearly identify origin of Chrysanthemi Indici Flos, these samples (C. indicum, C. morifolium, C. zawadskii var. latilobum and A. spathulifolius) were analyzed from five barcoding regions of chloroplast DNA (rbcL, matK, rpoB, atpF-atpH) and nuclear ribosomal DNA (ITS2) to evaluate the ability of discrimination for each barcoding region. Based on genetic distance, the percent of variable sites were provided the highest ITS2 value (56.9%), followed by atpF-atpH (48.18%), matK (27.2%), psbK (8.2%), and rbcL (2.9%). Comparative analysis based on the complete genome sequence of the petL-petG region INDEL (insertion/deletion) that the gene annotations were registered to the GenBank (accession number: JN-867592.1, NC-020092.1, MF-034027.1, NF-279514.1). Conclusion : From the above results, we may suggest that the petL-petG region INDEL analysis were conducted for molecular authentication of four plants (C. indicum, C. morifolium, C. zawadskii var. latilobum and A. spathulifolius). The findings of results indicated that petL-petG region might be established INDEL analysis systems and hence were proved to be an effective tools for molecular evaluation and comparison of “Chrysanthemi Indici Flos” with other plants.

Background : Agastache rugosa (A. rugosa), belonging to the Lamiaceae family, is a medicinal plant mainly distributed in Korea and contains various phenolic compounds revealing anti-fungal and anti-HIV properties. This study is aim to investigate change in phenylpropanoid content of flowers at different developmental stages using high performance liquid chromatography (HPLC) and quantitative real time polymerase chain reaction (qRT-PCR). Methods and Results : The variation in the transcriptional level of phenylpropanoid biosynthetic genes and phenylpropanoid contents in the flowers of A. rugosa at different developmental stages was analyzed. The transcript levels of phenylpropanoid biosynthesis genes, including ArPAL (phenylalanine ammonia-lyase), ArC4H (cinnamate 4-hydroxylase), and ArCHS (Chalcone synthase), were high in flowers at 1st stage compared with flowers at 2nd and 3rd stages. On the other hand, the expression levels of flavonoid biosynthesis genes, including ArTAT (tyrosine amino transferase), ArHPPR (hydroxyl phenylpyruvate reductase), and ArRAS (rosmarinic acid synthase), were higher in flowers at 3rd stage than those of flowers at 1st and 2nd. These results were consistent with HPLC analysis revealing that most phenolic compounds were higher in flowers at 1st and 2nd stage but the level of rosmarinic acid was the highest in 3rd stage. Conclusion : Our findings provide the information on change in phenylpropanoid biosynthesis in A. rugosa flowers at different developmental stages.

Background : Lycoris radiata belongs to the Amaryllidaceae family and is a bulbous plant native to South Korea, China, and Japan. Galantamine, a representative alkaloid of Amaryllidaceae plants, including L. radiata, exhibits selective and dominant acetylcholinesterase inhibition. In this study, transcriptome analysis of L. radiata was performed. Methods and Results : Genes for galantamine biosynthesis were used to design primers for qRT-PCR. Quantitative RT-PCR was performed with LrActin as a reference gene for normalization. The RT-PCR results reveal the expression of LrPAL A and LrC4H at an early stage in the pathway. Interestingly, the expression of these genes was significantly higher in roots. However, the expression levels of LrNNR and LrN4OMT, which are closely involved in galantamine biosynthesis, were significantly higher in bulbs than leaves and roots. The expression levels of LrPAL B, LrTYDC, LrCYP98A3 and LrCYP76T were not significantly different among the different parts of the plants tested. HPLC analysis confirmed the presence of galantamine in all the organs, including the root (0.53 ± 0.07 ㎎/g dry weight), bulb (0.27 ± 0.04 ㎎/g dry weight), and leaf (0.75 ± 0.09 ㎎/g dry weight). The galantamine level in the bulb was 1.42 and 2.78 times higher than that in the root and leaf, respectively. The results of qRT-PCR for the eight galantamine genes revealed relatively high levels of genes expressed early, including LrPAL A, LrPAL B, LrC4H, and LrTYDC in the roots. However, in the bulbs, the levels of LrNNR and LrN4OMT were higher, which are crucial for galantamine biosynthesis. It also explains why bulbs contain high amounts of galantamine, which is likely due to the increased expression of LrNNR and LrN4OMT and the high levels of LrCYP96T, although the genes expressed early were expressed at high levels in the root. Conclusion : Transcript data of plants grown in a growth chamber revealed high expression levels of LrNNR and LrN4OMT genes that are closely involved in galantamine biosynthesis, and, as expected, we observed higher amounts of galantamine in the bulbs than in the root and leaves.

Background : Tropospheric ozone (O3) is a secondary air pollutant that negatively affects numerous agricultural crop and forest. The tropospheric ozone is constantly increasing due to fossil fuel air pollutants. Here, we study the response of tartary buckwheat to ozone gas includes physiological and biochemical changes such as change in gene expression and metabolism. Methods and Results : Tartary buckwheat plants have green stems and leaves under normal conditions, while the plants exposed to the ozone have red stems and reddish green leaves. The expression of most flavonoid biosynthetic genes were significantly upregulated in ozone-treated buckwheat plants, exceting the expression of FtF3’H2. The contents of two anthocyanins, cyanidin 3-O-glucoside and cyanidin 3-O-rutinoside, were significantly increased by ozone treatment. From the metabolic profiling based on the GC-TOF-MS analysis, we identified the effect of ozone on thirty-five metabolites, including sugars, amino acids, and organic acids. Most of the metabolites result in significantly decreased or nearly remain unchanged in the ozone-treated plants compared with untreated plants, excepting alanine, proline, tryptophan, sucrose, and raffinose. To identify the effect of ozone on the leaf, we analyzed the epidermal cells on the leaf surface by scanning electron microscopy. Interestingly, amount of epidermal cells were partially destructed in ozone-treated plants. Conclusion : By analyzing both primary and secondary metabolites of tartary buchwheat without or with ozone, we identified that ozone affects the modulation of the metabolites as well as gene expression in tartary buchwheat.

Background : Miscanthus sinensis is a diploid hybrid and a temperate, perennial, cross-pollinating grass used as bioenergy plant, biomass production and high quality cellulose and ethanol production. This study was to carried out to investigate the expression of MsCOMT gene and the variation of lignocellulosic component and phenolic compounds contents in transgenic plants. Methods and Results : Multiple bands were detected from the homologous region of the COMT gene in PCR analysis. In order to obtain more detailed results, putative transgenic lines were estimated by RT-PCR analysis to confirm the expression of mRNA. Also, analysis of the lignin, cellulose, hemicellulose, and phenolic compound contents of transgenic Miscanthus plants were performed. Total lignin content of transgenic plants was lower than that of the control plant due to reduced caffeic acid O-methyltransferase (COMT) gene expression related to lignin production. Cellulose and hemicellulose contents in transgenic plants were not increased. Variation in cellulose and hemicellulose contents had no correlation with variation in lignin content of transgenic plants. Conclusion : In conclusion, transgenic M. sinensis was obtained with down-regulated COMT gene. Lignin synthesis was decreased what offers possibility of crop modification for facilitated biofuel production.

It is important for radish to have late flowering characteristics especially in the case of spring and winter cultivars. To understand late flowering characteristics of radish at the molecular level the flowering time genes of two radish lines (NH-JS1 and NH-JS2) with different flowering time were compared by re-sequencing their genomes. There were a total of 872,587 SNPs and 194,637 INDELs between the two lines. The SNP density of each chromosome was relatively uniform throughout, but the region with low SNP density was found at the end of R3 and the middle of R9. To compare the flowering time genes of the two lines, we first looked for the flowering time genes in radish using Arabidopsis thaliana flowering time genes. As a result, homologs of radish were found for most flowering time genes, but FRIGIDA was not found. Among 224 radish flowering time gene-homologs found, 97 genes showed more than one sequence difference (SNP or INDEL) between the two lines, and 127 genes had no difference. In particular, no sequence differences were found in FT, CO, and FLC, core flowering time control genes. Rs350520 (FVE), Rs193800 (CURLY LEAF) and Rs255320 (ATX1) with more than 100 sequence variations were expected to have a significant effect on flowering time difference between the two lines. These results will be of great help in understanding the flowering timing difference between the two lines at the molecular level.

Background : Although Oplopana elatus is a valuable medicinal plant that, like ginseng, recommends itself as source of herbal preparations, little molecular information on this plant is available. Therefore, to analyze the terpenoid biosynthetic pathway in O. elatus, we employed an RNA-seq approach. Methods and Results : To generate a transcriptome library of O. elatus, total RNA was extracted from different tissues, including leaves, stem, and root. A pooled RNA sample was prepared by combining equivalent RNA from different tissues. Using pair-end read with the Illumina platform, a total of 78,646,554 raw sequencing reads were generated. After data cleaning, approximately 77 million high-quality reads were obtained with 98% G20 (base quality of greater than 20). The high-quality reads were assembled into 208,959 unigenes with an average length of 1,073 bp and an N50 of 1,768 bp. Consistent with our prediction, which was based on the KEGG pathway assignment, we found 122 unigenes encoding 47 putative enzymes involved in the biosynthesis of terpenoid and triterpenoid saponins in the O. elatus transcriptome library. Conclusion : The transcriptome dataset generated in this study will serve as a valuable resource for accelerating genomic and functional genomic research in O. elatus and in the family Araliaceae.

Background : Cudrania tricuspidata Bureau is a widely used medicinal perennial woody plant. Obtaining information about the genetic diversity of plant populations is highly important for conservation and germplasm utilization. In this study, we developed single nucleotide polymorphism (SNP) markers derived from chloroplast genomic sequences to identify distinct Korean-specific ecotypes of C. tricuspidata via amplification refractory mutation system (ARMS)-PCR analyses. We performed molecular authentication of twelve C. tricuspidata ecotypes from different regions using DNA sequences in the chloroplast TrnL-F intergenic region. Methods and Results : SNPs were identified based on the results of nucleotide sequence for the intergenic region of TrnL-TrnF gene (chloroplast). Molecular markers were designed for those SNPs with additional mutations on the second base from SNPs for amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). HRM pattern analyses were performed using the Mx3005P QPCR System (Agilent Technologies, CA, USA). Conclusion : We collected 12 individual lines of C. tricuspidata from various region in South Korea and China. Based on the nucleotide sequence in the trnL-trnF intergenic region of these lines, six SNPs and a deletion of 12 bps were identified and 12 individual lines were able to be grouped in one Korean ecotype and two different ecotypes of chinese lines, chinese line 1 and 2. The SNP markers developed in this study are useful for rapidly identifying these specific C. tricuspidata ecotypes collected from different regions.

Background : Momordica charantia L. (M. charantia) is a member of the family Cucurbitaceae, used as a medicine herb in traditional medicine. In this study, we present the sequencing, de novo assembly and analysis of the transcriptome of M. charantia and provide a global description of relationship between putative phenylpropanoid and flavonoid biosynthesis genes and alteration of phenylpropanoid and flavonoid content during different organs and plantlet of M. charantia. Methods and Results : The transcriptome of M. charantia was constructed by using an Illumina Nexteseq500 sequencing system. Out of 68,073,862 total reads, approximately 88,703 unigenes were identified with a length of 898 bp. Alternatively, transcriptomic data, 10cDNAs (McPAL, McC4H, Mc4CL, McCOMT, McCHS, McCHI, McF3H, McFLS, McDFR and Mc3GT) encoded phenylpropanoid and flavonoid biosynthetic genes. The expression levels and the accumulation of trans-cinnamic acid, benzoic acid, 4-hydroxyvbenzoic acid, p-coumaric acid, chlorogenic acid, caffeic acid, catechin hydrate, ferulic acid, and rutin were investigated in different organs and plantlets. Mainly, phenylpropanoids and flavonoids accumulated in leaves and flowers, whereas low levels accumulated in roots. Collectively, these results indicate that the putative McPAL, McC4H, McCOMT, McFLS, and Mc3GT might be key factors for controlling phenylpropanoid and flavonoid contents in M. charantia. Conclusion : In this study, we present the sequencing, de novo assembly and analysis of the transcriptome of M. charantia. We also compared gene expression and compound analysis of phenylpropanoid and flavonoid in different organs and plantlet of M. charantia. These results indicate that McPAL, McC4H, McCOMT, McFLS, and Mc3GT are key regulators of phenylpropanoid and flavonoid accumulation in M. charantia