에메리개미 (Vollenhovia emeryi Wheeler)의 날개이형체의 유전체 크기 추정

노푸름,박소연,최재천,정길상,Noh, Pureum,Park, Soyeon,Choe, Jae Chun,Jeong, Gilsang Korean Society of Applied Entomology 2018 한국응용곤충학회지 Vol.57 No.4

에메리개미는 여왕개미와 수개미가 유전적으로 복제되어 번식한다고 알려져 있으며, 여왕개미의 날개형태가 장시형과 단시형으로 나타난다. 장시형은 정상적인 날개형태이고, 이보다 짧은 날개형태는 단시형이라고 한다. 장시형과 단시형 모두 한 종으로 취급되지만, 두 가지 점에서 종지위에 대한 조사가 필요하다. 첫째, 자연 상태에서는 두 날개형이 함께 발견되지 않고, 둘째, 날개형이 육안으로 뚜렷하게 구분된다. 또한 복제되어 번식한 여왕개미가 단수체인지 배수체인지 조사가 필요하다. 따라서 우리는 본 연구에서 에메리개미 유전체 크기를 추정하여 두 날개형은 동종이며, 여왕개미는 배수체임을 확인하였다. In Vollenhovia emeryi (Hymenoptera: Myrmicinae), the queen and the male are known to be clonally reproduced. Its colonies can be classified into the two morphs with the wing length of the queen caste. The morph with normal wings is called the long-winged and the other the short-winged that is brachypterous. Even though the two morphs are considered a species, investigation on the species status of the two morphs was suggested with natural separation in nature and the distinctive wing morphology. It has yet to be determined whether the clonally reproduced queen caste is haploid or diploid. Our data clearly show that the two morphs are the same species and the queen caste is diploid on the basis of the genome size data comparison.

범용성 DNA 바코드 분석 기반 한국산 천남성속(Arisaema) 식물의 분자계통학적 연구

노푸름,한경숙,김욱진,양선규,최고야,고성철,문병철 한국자원식물학회 2018 한국자원식물학회지 Vol.31 No.1

국내에 분포하는 천남성속의 계통학적 유연관계와 한약재천남성(Arisaematis Rhizoma)으로 사용되는 천남성 3종(둥근잎천남성, 두루미천남성, 일파산남성)에 대한 분류학적 특징을분석하기 위하여 천남성속 식물에 대한 분자계통학적 연구를수행하였다. 3개의 범용성 DNA 바코드(ITS, matK, rbcL) 염기서열을 이용하여 국내 분포 천남성속 8분류군과 중국에 분포하는 약전수재 종 1분류군을 포함하는 9종 50개 시료와 같은 과의Dracunculus vulgaris를 군외군으로 하여 유연관계를 분석하였다. 3개의 개별 DNA 바코드 염기서열과 이들을 유합한 염기서열로 계통학적 유연관계를 분석한 결과, 천남성속의 9 분류군은 6개의 독립적인 분계조를 형성하며 구별되었으며(Clade I, 둥근잎천남성 및 천남성; Clade II, 점박이천남성 및 섬남성; Clade III, 큰천남성; Clade IV, 일파산남성; Clade V, 두루미천남성; Clade VI, 무늬천남성 및 거문천남성), 이들 6개의 분계조는 각각 Pedatisecta절, Sinarisaema절, 및 Tortuosa절로 분류되었다. 또한 이들 DNA 바코드 구간의 비교 결과는 천남성속 식물의 종 및 종 이하 분류 단위의 분류학적 재검토의 필요성에 대한 중요한 정보를 제공하였다. 하지만 대한민국약전에 수재되어 한약재로 사용가능한 3종의 천남성 기원종의 종내 분류학적연관성이나 분자계통학적 특징은 확인되지 않았다. Molecular phylogenetic analysis was conducted to evaluate the taxonomic relationships of genus Arisaema L. distributed in Korea and the molecular phylogenetic characteristics of three authentic Arisaema species for the herbal medicine Arisaematis Rhizoma (the rhizomes of A. amurense, A. heterophyllum, and A. erubescens). The sequences of three DNA barcodes (rDNA-ITS, matK, and rbcL) were analyzed using 50 samples of nine taxa consisted of eight Korean and one Chinese Arisaema with one outgroup (Dracunculus vulgaris). Both individual and combined phylogenetic analyses of three DNA barcode sequences revealed that the treated nine taxa are independently classified into six distinct clades (Clade I, A. amurense f. amurense and A. amurense f. serratum; Clade II, A. serratum and A. takesimense; Clade III, A. ringens; Clade IV, A. erubescens; Clade V, A. heterophyllum; Clade VI, A. thunbergii subsp. thunbergii and A. thunbergii subsp. geomundoense). These six clades were reasonably divided into three individual sections, Pedatisecta, Sinarisaema, and Tortuosa. Futhermore, the results of comparative DNA barcode sequences analyses provided a significant information for the taxonomic reconsideration of Arisaema L. at the specific and intraspecific level. However, we could not confirm the taxonomic characteristics or identity among the three authentic medicinal species through the molecular phylogenetic analyses of genus Arisaema L. for Arisaematis Rhizoma. Key words - Arisaema, DNA barcode, matK, Molecular phylogenetic analysis, rDNA-ITS, rbcL

DNA 바코드 분석을 통한 개자 및 운대자 감별

노푸름,여상민,김욱진,양선규,박인규,문병철 한국원예학회 2018 원예과학기술지 Vol.36 No.6

Brassica juncea and Brassica napus are economically important crops used for oilseed and foragewho’s seed is also used for herbal medicines, namely Brassicae Semen and Brassicae CampestrisSemen, respectively. Morphological discrimination of these two herbal medicines is difficultbecause the seeds of the Brassica species have similar shapes. Since the descriptions of the differentBrassicae Semen in the Korean and Chinese pharmacopoeia can be confused, it is necessary todevelop an accurate method for identification of the different species. Therefore, DNA barcodes ofsix Brassica species, B. juncea, B. napus, B. nigra, B. oleracea, B. rapa, and Sinapis alba, collectedfrom various locations and markets, were analyzed to develop a molecular marker for distinguishingBrassicae Semen and Brassicae Campestris Semen. Nucleotide sequence comparison and phylogeneticanalysis of internal transcribed spacer (ITS) regions based on the amphidiploid types and U'striangle provide useful genetic information for the identification of these two herbal medicines andBrassica species. However, the sequences of the chloroplast maturase K (matK) gene showedspecies-specific insertions and deletions or substitutions only for B. nigra, B. oleracea, and S. albabut not for B. juncea, B. napus, and B. rapa. The use of genome-specific marker nucleotides in ITSregions provide an objective and accurate method for authentication of Brassica species andvaluable information for further molecular marker development. 배추속(Brassica)에 속하는 갓과 유채는 유지 및 사료작물 등 다방면으로 이용되며 그 종자는 개자와 운대자라는 한약재로사용되는 경제적으로 중요한 작물이다. 배추속 식물의 종자는 형태적으로 유사하여 개자와 운대자의 경우 형태적 종 감별에 어려움이 있다. 또한 우리나라 대한약전과 중국약전에서 개자에 대한 규정에 차이가 있어 이로 인하여 개자와 백개자가 혼용될우려가 있기 때문에 정확한 동정방법이 필요한 현실이다. 본 연구는 국내·외 다양한 지역에서 수집되거나 현재 유통되고 있는갓(B. juncea), 유채(B. napus), 흑갓(B. nigra), 양배추(B. oleracea), 배추(B. rapa), 그리고 백겨자(Sinapis alba)를 분석하여개자 및 운대자의 종을 구별할 수 있는 유전자 마커를 개발하기 위해 수행하였다. ‘우장춘의 삼각형’에 따라 복이배체 type을확인할 수 있는 ITS 부위의 염기서열 비교와 계통 분석을 통하여 ITS 부위가 두 약재의 감별 및 배추속의 종 동정에 유용한 유전적 정보를 제공한다는 것을 확인하였다. 하지만 matK 유전자의 경우에는 흑갓, 양배추, 백겨자 3종을 구분할 수 있는 염기치환은 존재하였으나, 갓, 유채, 배추에서는 종 특이적인 염기삽입/결실 및 치환이 나타나지 않았다. 따라서 ITS 부위의 각 genome특이적인 marker nucleotide를 이용하여 개자 및 운대자의 객관적이고 정확한 종 동정이 가능하며, 향후 분자마커 개발에 유용한 정보로 활용할 수 있을 것으로 기대된다.

돌 틈에 핀 꽃처럼

노푸름 샘터사 2014 월간 샘터 Vol.530 No.-

특허 증액배상제도에 관한 고찰

노푸름 ( Nohpooreum ) 연세대학교 법학연구원 의료·과학기술과 법센터 2019 연세 의료·과학기술과 법 Vol.10 No.1

There is a myriad of inventions in today's world. The ocean of patent has brought much convenience to our life, but at the same time there are long-pending and boring patent litigations, of which the results could potentially change an entire industry. A recent example is the patent war between Samsung and Apple. While the need and the importance of patent are growing, the term “Patent troll” has been coined to refer to companies that buy up patents. In fact, although patent system has been around for decades, impacting our everyday lives, the existing patent laws have often been an unavailable remedy for plaintiffs. Some individuals or small and medium-sized enterprises(SMEs) simply cannot afford long-winded patent lawsuits. In addition, under current patent laws, there has been no difference between ‘damages for willful patent infringement’ and ‘damages for unintentional patent infringement'. To that end, there has even been a prevailing false perception that if an infringer would only have to pay reasonable royalty fee as damages when found liable for infringement, the infringer might be tempted to profit from patent infringement instead of paying for “a fair price” for a patent on the front end. In order to improve the existing system's such problems, an enhanced damages provision in patent law has been implemented since July 9th, 2019. According to the new provision, courts have the power to increase the damages up to three times the amount assessed as a lost profit if ‘willful infringement' was found to have occurred. While the purpose of the new provision is to provide a proper remedy for victims as well as to prevent patent infringement, as its foundation is from the U.S. patent laws, this thesis examines issues such as the problem of text interpretation or conflicts with the existing laws and suggests solutions for observed problems. In addition, this thesis explores the effects of the enhanced damages provision including the promotion of technology development and economic growth through protection of patent rights, activation of the patent market, and enhancement of competitiveness in the global patent landscape. The patent system requires continuous monitoring as well as assessment and provision from relevant authorities so that it does not frustrate its primary purpose. Further, bear in mind that one of the main purposes of the patent system is to contribute to the industrial development by fostering the use of inventions. Finally, the enhanced damages provision should minimize potential issues that may arise during the process of implementation and act as an element that does not hinder technology development due to excessive damages or excessive patent litigation.

한국에 서식하는 개미의 성비교란 공생체 감염 실태

노푸름,박소연,정길상,최재천 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10

국내에서 채집된 개미 24종 71개체를 대상으로, 세포 내에 공생하며 자신들의 증식을 위해 숙주의 성을 교란하는 성비교란 공생체인 Wolbachia, Spiroplasma, Cardinium, Rickettsia의 감염 여부를 판별하였다. 24종 중 Wolbachia에 감염되어 있는 종은 9종이었으며, 감염률은 37.5%로 네 종류의 성비교란 공생체 중 감염률이 가장 높았다. 채집된 종 중 스미스개미(Paratrechina flavipes), 에메리개미(Vollenhovia emeryi), 일본왕개미(Camponotus japonicus)는 채집 지역별로 Wolbachia에 감염되어있는 군체와 감염되어있지 않은 군체가 나뉘었다. 또한, 곰개미(Formica japonica)와 극동혹개미(Pheidole fervida)는 각각 11개 지역과 5개 지역에서 채집되었는데, 모든 채집 지역의 개체들이 Wolbachia에 감염되어있는 것으로 나타났다. Spiroplasma의 경우, 24종 중 코토쿠뿔개미(Myrmica kotokui) 단 한 종에서만 감염되어 있는 것을 확인하였다. Cardinium과 Rickettsia는 채집된 전체 개미 개체 모두에서 비감염인 것으로 확인되었다.

통초(通草), 목통(木通) 신속 감별용 ITS 염기서열 기반 SCAR 마커 및 Multiplex-SCAR 분석법 개발

노푸름 ( Pureum Noh ),김욱진 ( Wook Jin Kim ),박인규 ( Inkyu Park ),양선규 ( Sungyu Yang ),최고야 ( Goya Choi ),문병철 ( Byeong Cheol Moon ) 대한본초학회 2021 大韓本草學會誌 Vol.36 No.1

Objectives : Tetrapanacis Medulla and Akebiae Caulis are one of the most frequently adulterated herbal medicines because of their confusability of terms in the ancient writings and the similarity of morphological features of dried herbal products. The major adulterant is Aristolochia manshuriensis (Guanmutong) which has a serious safety concern with its toxicity. To ensure the safety and quality of the two herbal medicines, it is necessary to discriminate the toxic adulterant from authentic species. The aim of this study is to develop SCAR markers and to establish the multiplex-SCAR assay for discrimination of four plant species related to Tetrapanacis Medulla and Akebiae Caulis. Methods : ITS regions of fifteen samples of four species (Tetrapanax papyrifer , Fatsia japonica , Aristolochia manshuriensis, and Akebia quinata ) collected from different sites were amplified and sequenced. Fifteen obtained ITS sequences were aligned and analysed for the detection of species-specific sequence variations. The SCAR markers were designed based on the sequence alignments and then, multiplex-SCAR assay enhancing rapidity was optimized. Results : ITS sequences clearly distinguished the four species at the species level. The developed SCAR markers and multiplex-SCAR assay were successfully discriminated four species and detected the adulteration of commercial product samples by comparison of the amplified DNA fragment sizes. Conclusions : These SCAR markers and multiplex-SCAR assay are a rapid, simple, and reliable method to identify the authentic Tetrapanacis Medulla and Akebiae Caulis from adulterants. These genetic tools will be useful to ensure the safety and to standardize the quality of the two herbal medicines.

Population genetic structure based on mitochondrial DNA analysis of Ikonnikov’s whiskered bat (Myotis ikonnikovi—Chiroptera: Vespertilionidae) from Korea

박소연,노푸름,최유성,주성배,정길상,김선숙 한국생태학회 2019 Journal of Ecology and Environment Vol.43 No.4

Background: Ikonnikov’s whiskered bat (Myotis ikonnikovi) is found throughout the Korean Peninsula, as well as in Kazakhstan, Russia, Mongolia, China, and Japan. It is small-sized and primarily inhabits old-growth forests. The decrease and fragmentation of habitats due to increased human activity may influence the genetic structure of bat populations. This study was designed to elucidate the population genetic structure of M. ikonnikovi using mitochondrial genes (cytochrome oxidase I and cytochrome b). Results: The results showed that M. ikonnikovi populations from Korea have high genetic diversity. Although genetic differentiation was not detected for the COI gene, strong genetic differentiation of the Cytb gene between Mt. Jeombong and Mt. Jiri populations was observed. Moreover, the results indicated that the gene flow of the maternal lineage may be limited. Conclusions: This study is the first to identify the genetic population structure of M. ikonnikovi. We suggest that conservation of local populations is important for sustaining the genetic diversity of the bat, and comprehensive studies on factors causing habitat fragmentation are required.

ITS2 DNA 바코드 염기서열 분석을 통한 천속단과 산토끼꽃 감별

박인규,노푸름,김욱진,양선규,문병철 한약정보연구회 2017 한약정보연구회지 Vol.5 No.3

Dipsaci Radix was described as the roots of Dipsacus asperoides in the Korean pharmacopoeia, namely Sok-dan. In Korea, a herbaceous plant species Dipsacus japonicus belong to the same genus also distributed. Unfortunately, due to the morphological similarity of the areal parts and dried root, the accurate identification of these two plant is very difficult depending on the morphological features. Therefore, we introduced the genetic tools to distinguish D. asperoides and D. japonicus at the species levels based on the nuclear ribosomal RNA gene (nrDNA) internal transcribed spacer 2 (ITS2) region sequences. In comparison of ITS2 sequences using 7 and 2 samples of D. asperoides and D. japonicus, respectively, we identified seven distinct species-specific single nucleotide polymorphisms (SNPs), namely marker nucleotides, enough to discriminate both two plant species. Furthermore, these two species were clearly classified into distinct two group depending on species in the phylogenetic analysis of 7 Dipsacus species. These results indicate that the sequence variability and species-specific nucleotide substitutions of ITS2 region are useful genetic marker for identifying authentic Dispsaci Radix.

COI 염기서열 기반 백강잠 신속 감별용 SCAR marker 개발 - 백강잠 유전자 감별 -

김욱진,양선규,노푸름,박인규,최고야,송준호,문병철 대한본초학회 2019 大韓本草學會誌 Vol.34 No.5

Objectives : To ensure the safety, quality and pharmacological efficacy of Batryticatus Bombyx, it is important to discriminate with adulterants. In Korean Herbal Pharmacopoeias (KHP), the authentic species of Batryticatus Bombyx is defined only Bombyx mori. Therefore, the aim of this study is establishment of PCR assay method using the sequence characterized amplified region (SCAR) marker based on COI DNA barcode for discriminating six species related to Batryticatus Bombyx. Methods : Seventeen samples of six species (Bombyx mori, Bombyx mandarina, Rhodinia fugax, Oberthueria caeca, Actias artemis, and Caligula japponica) were collected from different habitate and nucleotide sequences of cytochrome c oxidase subunit I(COI) barcode regions were analyzed by Sanger sequencing methods. To develop SCAR-based PCR assay method, we designed species-specific primers based on COI sequence variabilities and verified those specificities using 17 samples of six species as well as commercial herbal medicines. Results : In comparative multiple analysis of COI sequences, six species were distinguished by species-specific nucleotides at the species level. To develop rapid and reliable PCR assay method for genetic authentication of Batryticatus Bombyx, therefore, we designed species-specific SCAR primers based on these nucleotide sequences and confirmed those specificities. Using these SCAR primers, We also established simple conventional PCR assay method using these SCAR primers at the species level. Conclusions : The comparative analysis of COI sequences and SCAR-based PCR assay methods represented equal results for distinguishing authentic Batryticatus Bombyx and adulterations at the species level. Therefore, our results are expected protecting adulteration of herbal medicine Batryticatus Bombyx.