본 연구는 국내 유통 방풍(S. divaricata, P. japonicum, G. littoralis) 한약재의 원산지 판별을 위해 수행 되었다. 이를 위 해, 형태적 특징비교 뿐만 아니라 엽록체(nrDNA-ITS2) 및 핵 DNA 바코드 유전자(cpDNA-matK, psbA-trnH, rpoB2와 rpoC1)를 이용한 염기서열 분석을 수행하였다. 방풍류 3종의 형태적 특징을 비교한 결과, 잎 모양과 거치의 형태에서 가장 큰 차이를 보인 반면, 건조약재의 경우 육안으로 구별하기에 어려움이 있었다. DNA 수준에서의 차이를 비교하기 위해 DNA 바코드 후보 유전자들을 이용한 방풍류 3종의 식물자원 에 대한 염기서열 분석을 수행한 후 판별 마커 개발 가능성이 있는 ITS2와 matK, psbA-trnH 세 primer를 선발하였다. 이 를 국내 유통 한약재에 적용한 경우 방풍은 S. divaricata와, 식방풍은 P. japonicum과, 해방풍은 G. littoralis와 동일한 것 으로 확인되었다. 중국산 방풍은 S. divaricata와 동일종으로 바르게 표기되어 유통되나, 국산 방풍은 P. japonicum과 동일 종으로 식방풍(갯기름나물)의 뿌리를 건조시켜 방풍으로 혼용 유통됨을 확인하였다. ITS2 구간의 염기서열 분석 결과, 식방 풍의 경우 두 그룹으로 나눠져 동일 종 내에 유전적 변이가 있음이 확인되었다. 따라서 본 연구결과 방풍류 3종 판별을 위 해 nrDNA-ITS2와 cpDNA-matK, psbA-trnH의 사용이 유용 할 것이며, 차후 방풍류 한약재의 판별을 위한 마커 개발 연 구의 기초 자료로서 활용될 수 있을 것으로 사료된다.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

A xylan-decomposing Gram-positive bacterium, Cellulosimicrobium cellulans DY-8, was isolated from the gut of a wood-feeding longicorn beetle, Moechotypa diphysis. To amplify a partial fragment of the GH 10 β-1,4- xylanase (XylC) gene of strain DY-8, two degenerated oligonucleotide primers were designed based on strictly conserved regions (WDVVNE and ITELLDV) in the GH family 10 xylanolytic enzymes. The full gene (1488-bp) of XylC, which was predicted to encode a protein consisting of 495 amino acids with a molecular mass of 52.0 kDa and a calculated pI of 6.49, was cloned by repeated DNA walking and nested PCR protocols. The results of a protein blast survey showed that XylC was a β -1,4-xylanase comprised of an N-terminal catalytic GH10 domain (from Ser48 to Leu338) and a C-terminal RICIN domain (from Tyr359 to Leu492). This overall structure of XylC was 57% identical to that of Actinoplanes sp. SE50/110 β -1,4-xylanase (Accession number: YP_006265966), which has not yet been biochemically characterized.

Sacbood virus (SBV) is an infectious disease, resulting in failure to pupate and death and kSBV is a disease caused perish Apis cerana of 75% in South Korea. RNA dependent RNA Polymerase(RdRP) is one of polyprotein of viral genome and an enzyme that catalyzes the replication of RNA from an RNA templates and an essential protein encoded in the genomes of all RNA containing viruses with no DNA stages. In this study, recombinant construct with RdRP partial region of kSBV was used for sequence analysis to clarify about Korean SBV. As a result it could be determined that the virus develops by infection of Korean Apis cerana called kSBV. Also, we named Apis cerana-kSBV-region to the name of the unique region of gene that kSBV has. In comparison of the RdRP region of bee RNA virus on nucleotide sequence, its sequence from same species have less variability as well as each virus species has a certainty of RdRp region. It indicated that mutations of RdRP region of each virus species is able to be a useful indicator of honeybee virus detection.

The purpose of this study was to identify fast and effectively the Nosema disease of bumble bees from Gangwon province in Korea. Bumble bees are crucial pollinators of various crops and microsporidia are the critical infections of these hosts. The origin of the bumble bees is probably being known from the Britain. Also, the following species of bumble bees have been used around the world: Bombus terrestris, Bombus lucorum, Bombus occidentalis, Bombus ignites and Bombus impatiens. When bumble bees are infected with N. bombi, their abdomens can become distended, paralyzed and infected workers and they often become sluggish and die early. We have identified the morphology of the microsporidium by light and electron microscopy and have found that the morphology of the microsporidium is rounded spore morph, with fairly small spores as described before in many other articles. For the specific and sensitive diagnosis of the microsporidian parasite N. bombi in bumble bees, we have developed the improved method of polymerase chain reaction (PCR) conditions for expeditious diagnosis. Two pairs of primers were tested on N. bombi and the related microsporidia Nosema apis and Nosema sp., both of which infect bumble bees and honey bees and further we have verified and analyzed DNA sequence data of N. bombi in bumble bees by using the BLAST server at the National Center for Biotechnology Information.

The effect of a sputter deposition sequence of Cu, Zn, and Sn metal layers on the properties of Cu2ZnSnS4 (CZTS) was systematically studied for solar cell applications. The set of Cu/Sn/Zn/Cu multi metal films was deposited on a Mo/SiO2/Si wafer using dc sputtering. CZTS films were prepared through a sulfurization process of the Cu/Sn/Zn/Cu metal layers at 500˚C in a H2S gas environment. H2S (0.1%) gas of 200 standard cubic centimeters per minute was supplied in the cold-wall sulfurization reactor. The metal film prepared by one-cycle deposition of Cu(360 nm)/Sn(400 nm)/Zn(400 nm)/Cu(440 nm) had a relatively rough surface due to a well-developed columnar structure growth. A dense and smooth metal surface was achieved for two- or three-cycle deposition of Cu/Sn/Zn/Cu, in which each metal layer thickness was decreased to 200 nm. Moreover, the three-cycle deposition sample showed the best CZTS kesterite structures after 5 hr sulfurization treatment. The two- and three-cycle Cu/Sn/Zn/Cu samples showed high-efficient photoluminescence (PL) spectra after a 3 hr sulfurization treatment, wheres the one-cycle sample yielded poor PL efficiency. The PL spectra of the three-cycle sample showed a broad peak in the range of 700-1000 nm, peaked at 870 nm (1.425 eV). This result is in good agreement with the reported bandgap energy of CZTS.

본 논문에서는 도심지 지하에 터널 전력구를 건설하는 경우 시공단계별 영향을 고려한 구조해석을 수행하였다. 해석대상의 도심지 지하에는 여러 종류의 다양한 라이프라인 구조체가 설치되어 있다. 터널전력구의 구조해석에는 지반체의 유한요소해석 프로그램인 MPDAP을 사용하였다. 라이프라인 구조체와 터널 전력구 사이의 이격거리가 가장 작은 대표적인 3개의 단면에 대하여 구조해석을 수행하였다. 터널의 굴착단계별 유한요소해석에서 발생되는 평형불균형성 문제는 평형섭동개념을 적용하여 해결하였다. 또한 터널 굴착에 의한 시간의존 변형의 영향은 하중분담율을 사용하여 시공단계별로 고려하였다. 본 연구에서 검토한 3개의 대표단면에서는 터널 전력구 주변 지반체에서 발생하는 최대변위값은 허용변위값이내를 보여주었다.

Worldwide studies on Apis cerana variation for biogeography and genetic diversity depended largely on a 86~93 bp-long mitochondrial non-coding region (internal spacer region) located between tRNALeu and COII (named as NC2), possibly due to higher variability among available markers. In order to incorporate the A. cerana occurring in South Korea into world extensive data, we also sequenced the NC2 from 118 A. cerana samples collected over nine Korean localities and 66 A. cerana samples over seven Asian localities, such as China, Vietnam, and Thailand. These data were combined with preexisting world data to scrutinize genetic relationships of A. cerana in South Korea to outside distributional range. Sequencing of 184 samples provided a total of ten haplotypes: five from Korea, six from China, one from Vietnam, and two from Thailand. Among them eight were new, whereas two were previously reported ones. Phylogenetic analysis of A. cerana NC2 haplotypes so far found including ours has confirmed the presence of four major groups of A. cerana (Asian mainland group, Sundaland group, Palawan group, and Luzon-Mindahnao group) and all haplotypes found in this study also were included in the Asian mainland group. In order to find further variable regions that can be used as sequence-based marker several mitochondrial non-coding regions and nuclear intron regions are in the middle of testing.

In the current Internet environment, a lot of multimedia information is used; such as text, sound, image, and video. Among those information video is the most valuable and meaningful data. Therefore, the key frames of a video are very important role to recognize the whole sequence contents. In this paper, we propose a new key frame extraction scheme using chi-square and color histogram. The proposed key frame extraction scheme showed the better results compared to the previous studies. The proposed scheme results will be applied to the many computer game programs for the fast scene detection and the recognition of the game contents.

비정형 초고층 건물의 계획 및 시공이 늘어남에 따라 이 연구에서는 프로토타입 모델에 대한 시공단계해석의 적용을 통하여 비정형 초고층 건물의 시공 중 구조적 거동을 분석하고자 하였다. 비틀림 초고층 건물을 대상으로 횡력저항시스템, 비틀림각도, 공법 조건에 따른 총 18개의 모델을 선정하였다. 횡력저항시스템으로는 다이아그리드 시스템과 가새튜브 시스템을 적용하였으며, 각 횡력저항시스템별로 0〫, 1〫, 2〫 비틀림각도를 갖는 세 가지 평면 형태와 외곽 튜브와 내부 골조의 시공순서에 따른 세 가지 공법을 가정하였다. 시공 중인 초고층 건물의 구조적 성능은 시공단계해석의 횡변위 결과를 통하여 분석되었으며, 골조 공기와 최대 양중량과 같은 시공성능이 함께 비교되었다.

본 연구는 국내 유통 감초약재의 원산지 판별을 위해 수행되었다. 이를 위해 약재로 쓰이는 만주감초(Glycyrrhiza uralensis) 및 유럽감초(G. glabra)와 약재로 사용되지 않는 국내 자생종 개감초(G. pallidiflora) 식물의 형태적 차이 및 엽록체와 핵 DNA 구간의 염기서열의 차이를 구명하였다. 감초식물 3종의 형태적 특징을 비교한 결과, 잎과 뿌리에서 차이를 보였다. 특히 잎의 형태에 있어서 중국감초와 유럽감초는난형 혹은 타원형인 반면, 개감초는 엽장/엽폭비가 큰 피침형이었다. 또한 건조된 뿌리로 시중되는 감초 한약재의 경우, 중국산과 한국산은 백색인 반면 우즈베키스탄산은 황백색이었다.감초식물 3종을 대상으로 DNA 수준에서의 차이를 비교한 결과 rpoB2, rpoC1에서는 G. uralensis와 G. glabra가 구분되지않았으며, psbA-trnH의 경우 단 한 구간의 SNP에서 차이를 보였다. 반면 ITS2에 의해 증폭된 450bp의 염기서열을 비교한 결과, G. glabra는 98, 99, 100번째 위치에서, G. pallidiflora는 137, 161, 164, 203, 296위치에서 감초 종간판별을 위한 특이적 SNP가 확인되었다. 또한, phylogenetic tree 분석을 통해, 약재로 쓰이는 G. uralensis과 G. glabra는 약재로 쓰이지 않는 G. pallidiflora에 비하여 유전적 거리가 가까움을 확인하였다. 본 표준시료에서 얻은 결과를 유통 한약재 원산지 판별에 적용한 결과, 중국산과 한국산의 한약재는 G. uralensis와 염기서열이 일치하였고, 우즈베키스탄으로부터 수입된 한약재는 G. glabra와 염기서열이 일치됨을 확인하였다. 이에 감초의 생체 및 한약재의 원산지 판별을 위해 ITS2를 이용한 분자수준에서의 판별이 유용하다고 판단된다.

콩과(Fabaceae) 작물 해충인 팥나방(Matsumuraeses phaseoli)과 어리팥나방(M. falcana) (나비목: 잎말이나방과)은 형태적으로 매우 유사하여 종 구별이 힘든 것으로 알려져 있다. 본 연구에서는 PCR-SSP(PCR with Sequence Specific Primers) 방법으로 두 종을 빠르고 정확하게 구별할 수 있는 판별법을 찾고자 두 종의 미토콘드리아 시토크롬 옥시다제 I(mtCOI) DNA 부분영역(439 bp)의 염기서열을 해독하였다. 그리고 다른 나방 종의 mtCOI 염기서열과 함께 나열하여 비교한 후 팥나방과 어리팥나방에서 종 특이적으로 차이가 나는 단일 뉴클레오티드를 프라이머의 3ʹ말단으로 하는 염기서열 특이 프라이머 조합을 만들었다. PCR 산물들을 전기영동 한 결과, 어리팥나방은 245 bp, 팥나방은 409 bp와 245 bp의 특이적 밴드 패턴을 보여 두 종을 구별할 수 있었다.

Honeybees (Apis mellifera) adapted themselves to different geographical and climatical conditions since they had been introduced in Korea. Beekeepers have tried to breed valuable lineages with artificial insemination or conventional mating techniques. However, evaluation of breeding resultants still relies on timeconsuming observation data. Genetical characterization of breeds has proven its usefulness to preserve genetic resources of livestock. In recent years, microsatellites are most commonly used to evaluate population structures and diversities of living organisms in that the characteristics of locus specificity, rich polymorphism, abundant and random distribution over the genome, and their co-dominant inheritance. Determining classic genetic distances using neutral, highly polymorphic microsatellite markers is a reliable method to investigate genetic relationships and breed differentiation. This methodology can be used to establish preservation priorities for livestock breeds. The final aim of this study was to develop potent markers for assessing genetic structure of lineages after artificial insemination. In this study, the genetic structure of ten microsatellite markers were sequenced or analysed with polymerase chain reaction for eleven European honeybee populations. The results may help to develop reliable microsatellite markers for more efficient preservation strategies of valuable honeybee breeds.

황해 경기만은 제4기에 반복된 해침과 해퇴로 4개의 해침-해퇴 퇴적체(DS-1, DS-2, DS-3, DS-4)를 형성하였다. 본 연구는 황해 경기만 퇴적체의 형성을 6개 퇴적단계로 나누어 설명한다. A 단계는 MIS-6 저해수면 시기로서 큰 규모의 해수면 하강으로 인해 대부분의 지역이 대기에 노출되고 광범위한 하도 침식 및 풍화작용의 영향을 받았다. 이어지는 B 단계는 MIS-5e 까지 빠른 해수면 상승과정에서 MIS-5 시퀀스의 하부 해침퇴적체가 형성되었고, 다음에 이어지는 MIS-5d부터 MIS-4 저해수면 시기까지의 C 단계에서는 MIS-5 시퀀스의 상부인 해퇴퇴적체가 만들어졌다. 다음의 D 단계는 MIS-4 저해수면 시기부터 MIS-3c 고해수면 시기까지로 하도 침식 및 하도 충전 구조로 이루어진 MIS-3 해침 퇴적체가 형성되었다. 다음의 E 단계에서는 LGM 시기까지 계속적인 해수면 하강이 있었고 외해쪽에서는 천해 기원의 MIS-3 해퇴퇴적체가 만들어진 반면에, 내륙쪽에서는 노출된 환경에서 하도 충전 퇴적체나 범람원 퇴적체가 형성되었다. 마지막 단계인 F 단계는 황해 전체적으로 홀로세 해침이 발생하였고, 이 시기에 외해쪽에서는 대륙붕 사질 퇴적체와 조석사주 퇴적체가 형성되어 고해수면 시기인 현재까지 퇴적이 일어나고 있다.

XIST has been known to long-non coding RNA which regulate X-chromosome inactivation in female mammal and the gene has been suggested to having important role in early embryo development and embryonic stem cell. However, its coding region has been unclear in pig. To determine the coding region of XIST in pig, we have examined candidate site of XIST coding region in pig by BLAST, PCR, and sequencing. By comparing pig whole genome sequence (Sus scrofa 10.2) with human, murine, and bovine XIST transcript sequence using BLAST, we selected candidate coding region of XIST in pig. The result showed XIST is coded on the minus strand of NW_003612825 contig and its length was nearly 32kb which was similar to the length of human and bovine XIST gene. With the candidate model, we performed RT-PCR to confirm the coding region of XIST with 24 primer pairs and they were expressed only female porcine embryonic fibroblast (PEF) but not in male PEF. By designing candidate intron spaning primer we could confirmed candidate intron is present between first and last exon (distance, 9.2kb vs product size, 2kb). The seqeucne of amplicon was analyzed and we could confirmed there were 5 small exons (less than 400 bp) like XIST coding region of other species which have 4 to 5 small exon between first and last exon. To confirm coding strand in pig, we conducted strand specific reverse transcription. We confirmed candidate XIST was coded on the negative strand of contig on X-chromosome as the result of homology analysis by BLAST. With the candidate pig XIST sequence, we aligned the sequence with XIST sequences of 3 species, human, mouse, and bull by clustalW. These result showed candidate sequence of pig XIST is most similar to that of bovine and the homology between pig and human was higher than result between mouse and human. These results could support for X chromosome inactivation analysis and the function of XIST in pig preimplantation embryos. * This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (No. 2012006276)

느타리버섯은 시중의 재배기술이 발전하여 느타리와 큰느타리의 전체 생산 및 소비가 버섯전체생산의 약 50%를 차지하여 재배품종의 주류를 형성하고 있다. 하지만 UPOV 및 FTA 등의 문호개방에 따라 로열티 지불 등에 대응할 수 있는 품종육성이나 신기술 개발이 시급한 실정이다. 본 연구에서는 느타리버섯의 성장단계별 유전적 발현을 분석하기 위해 cDNA 라이브러리를 성장단계별 10단계로 구분하여 12,342 ESTs를 구축하였다. 기존에 등록된 29,211 EST와 함께 분석한 결과 contig 4,939로 40%를 보여주었다. 이것을 기능별로 분류하였을 때 생물학적 과정 (Biological process) 27%, 세포 요소 (Cellular component) 35%, 분자 기능 (Molecular function) 38%로 분포하였다. 생물학적 과정에서는 생리학적 과정과 세포 과정이 각각 50%와 46%, 세포 요소에서는 세포와 세포부분이 각각 35%, 분자 기능에서는 촉매활성과 결합력이 각각 44%와 40%를 차지하였다. 유전자군별로 다양한 발현 패턴을 보여주었으며 조직특이적 발현은 약 13%를 나타내었다.

In the present study, a phylogenetic analysis was undertaken based on the internal transcribed spacer (ITS) rDNA and partial β-tubulin gene sequence of the Ganoderma species. The size of the ITS rDNA regions from different Ganoderma species varied from 625 to 673 bp, and those of the partial β-tubulin gene sequence were 419 bp. Based on the results, a phylogenetic tree was prepared which revealed that Korean Ganoderma lucidum strains belong in a single group along with a G. lucidum strain from Bangladesh.

In the transcriptome surveys of Laodelphax striatellus, several cDNA sequences showed a high level of similarities to the insect picorna-like virus genomes. Interestingly, there was no sequence similarity between picorna-like virus sequences from the RSV-viruliferous and those from the non-viruliferous L. striatellus. Picorna-like virus from the non-viruliferous L. striatellus was a geographical isolate of Himetobi P virus (HiPV). The genome of the HiPV was 9,272 nt in length excluding the poly(A) tail and contained two open reading frames (ORFs), which were separated by a 176 nt intergenic region that functions as an internal ribosome entry site (IRES). The 5' ORF encodes the non-structural proteins and the 3' ORF encodes the capsid proteins. The partial genomic RNA of the picorna-like virus from the RSV-viruliferous L. striatellus, LsPV-2, was 8,769 nt in length excluding the poly(A) tail and contained a single, large open reading frame (nt 1–8,535) encoding a 2,845 aa polyprotein. In terms of sequence similarity, identity, and genome organization, LsPV-2 resembled insect picornalike viruses belonging to the family Iflaviridae. A phylogenetic analysis based on RNA-dependent RNA polymerase (RdRp) sequence showed that LsPV-2 was most closely related to the deformed wing virus (DWV). The HiPV and LsPV-2 were incompatible each other in L. striatellus, suggesting that these two picorna-like viruses may have important functions in transmission of the RSV.