무포자형성 자연돌연변이 균주의 선발 및 특성검정 결과 ASI 2069 균주가 무포자이면서 수량성이 높으나 상품적 가치가 없어 원형질체 재생에 의한 단핵화로 Nh36 (neohaplont 36) 등 4균주를 분리하였다. 원형1호(ASI 2180) 및 무포자느타리간의 단핵교배 (Mon-Mon)를 시도한 결과 128교배조합수를 얻어 이 중 30균주를 특성 검정한 다음 자실체형성 및 포자비산량 조사에 의해 소포자형성 유망 13균주를 선발하였다. 무포자균주의 스크리닝법을 개발하기위해 포자형성 및 감수분열에 관여하는 helicase, DMC1(recombinase) 및 Spo11(topoisomerase II)유전자를 이용하여 PCR 증폭한 결과 단핵화균주 Nh36 및 교배체 2균주 중에서 Spo11유전자가 증폭되지않아 무포자 균주 스크리닝 방법으로 가능하리라 사료된다.

Insects or insect remains found in beer are one of major issues in consumer claim. Accurate estimation of inflow time isa critical factor for the settlement of such claims related with beer-contaminating insects but no reliable methods have been developed. In an attempt to establish a molecular marker-based diagnostic method, the degradation rates of 18S rRNA genes in the insectssoaked in 500 ml beer were investigated by quantitative real-time PCR (qPCR) over one month period at room temperature. Among the six insect species tested, the house fly (Musca domestica) and honey bee (Apis mellifera) revealed high correlations (r2=0.974-0.990) between the degradation of 18S rRNA gene and inflow time. In these insects, statistically significant distinction was possible between the samples stored in beer less than 14 days and more than 14 days. Other insects, including the fruit fly, common house mosquito, German cockroach and Indian meal moth, displayed poor correlations, which appeared attributed to the inefficient genomic DNA extraction likely due to small sample size or disintegration of body parts during storage in beer. With proper improvement in DNA extraction, this 18S rRNA-based diagnostic method would be applicable for estimating the inflow time of beer-contaminating insects.

In this study, genotyping was executed by using 11 microsatellite markers (BM1824, BM2113, ETH10, ETH225, ETH3, INRA23, SPS115, TGLA122, TGLA227, TGLA53, and TGLA126) for diversity of 214 Hanwoo cows in Hoengseong area. Each marker's size and number of allele, observed heterozygosity, expected total heterozygosity, and polymorphism information content were analyzed by 11 Microsatellite marker. The average of size range was detected from 150.9 to 174.9 in Hanwoo cows of Hoengseong. The number of average allele was 10.0 that is similar to the average of Kangwon Hanwoo, which is known as 10.5 in the previous report. The average were 0.751 for observed heterozygosity, 0.760 for expected total heterozygosity, 0.725 for polymorphism information content, respectively. These results were similar to previous studies in Kangwon Hanwoo, National Hanwoo and Korean Proven Bulls. This study is expected to contribute for genetic improvement of Hanwoo cows in Hoengseong as a popular brand.

现代汉语的被动句可分为有标志被动句与无标志被动句。其中无标志被 动句与null subject主题句，在形式方面结构非常相似。因此，它们的分类 及分析方法上有很多错误。为了补充形式上的解释，本稿采取认知语言学的 观点来解释被动句的制约条件，进而来区分无标志被动句与null subject主 题句。按词汇概念，被动句的‘相(aspect)’和无标志被动句的事件可分别分析 为下面的意义结构。[[[xACT]CAUSE[BECOME[y<STATE>]]]CAUSE[BECOME[y<AF FECTED>]]] → [[do'[∅<manner>(y)]]CAUSE[BECOME[y<AFFECT ED>]]]。这个结构说明的最大的特征就是被影响论项(而不是受事论项)的存 在。而加上‘相(aspect)’的特征的话，可以表现为[y<RES-STATE=∅[[do' [∅<manner>(y)]]CAUSE[BECOME[y<AFFECTED>]]]>]。如果句子符 合这些制约条件，它可以分确定为无标志被动句。

벼의 약 및 현미 배양효율과 관련된 DNA marker를 이용하여 인디카형 벼 품종인 'IR 36'의 조직배양 효율을 개선하기 위하여 실험한 결과를 요약하면 다음과 같다. 벼품종 간에 약 및 현미배양 효율을 비교한 결과 자포니카 > 통일형 > 인디카 형의 순으로 나타났다. 그러나 MGRI집단의 약배양에서 식물체분화율이 높은 계통으로 선발된 'MGRI 079'와 'MGRI 036'의 약배양 효율은 각각 19.8%, 19.9%로 가장 높게 나타났다. 'MGRI 079'에 'IR 36'이 여교배되어 양성된 90 계통에 대한 marker검정을 실시하여 positive band를 나타내는 34계통을 선발할 수 있었다. 선발된 34계통 중 10 계통의 약배양에서 캘러스 형성률은 'IR 36' 보다 현저히 높았다. 선발된 10 계통의 현미배양에서도 캘러스형성 능력과 식물체재분화율이 'IR 36' 보다 높게 나타났다. 계통 중에서 식물체분화능력이 높은 계통으로 선발된 -28은 간장이 'IR 36'보다 큰 편이었으나 출수기와 미립특성은 'IR 36'과 비슷하였다.

Two closely related species, the soybean podworm, Matsumuraeses phaseoli, and the podborer, M. falcana, gives differential economic damages on crops. It is difficult to discriminate these potential sympatric species by morphological characters. The goal of this study was to develop a discriminating molecular marker based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). A partial genomic fragment (500 bp) of mitochondrial cytochrome oxidaseⅠ (mtCOI) was sequenced in both species, in which restriction site by Rsa I was selected as a dichotomous marker. PCR-RFLP in the mtCOI region clearly discriminated both species.

Antibiotics marker-free with herbicide-resistant tall fescue plants were produced through Agrobacterium: mediated transformation system. Embryogenic calli derived calli were infected with Agrobacterium tumefaciens strain EHA105 harboring the pCAMBIA3300 vector containing the bar and the CP4-EPSPS genes. The PPT-resistant calli and plants were selected with 10 ㎎/L PPT, respectively. Soil-grown plants were obtained about 14-16 weeks after Agrobacterium-mediated transformation. Genetic transformation of the regenerated plants growing under selection was demonstrated by PCR, and Southern blot analysis revealed that one copies of the transgene were integrated into the plant genome of each transgenic plant. Expression of the bar and CP4-EPSPS genes in transgenic plants was confirmed by Northern blot analysis and CP4-EPSPS genes in transgenic plant ELISA uses antibody protein by ELISA Assays. Transgenic plants sprayed of two herbicides with glufosinate ammonium or glyphosate remained green and healthy. We therefore report here a successful and reliable Agrobacterium-mediated transformation of two herbicide-resistances and this method may be useful for routine transformation and has the potential to develop new varieties of tall fescue with several important genes.

Local and seasonal populations of the oriental fruit moth, Grapholita molesta , were monitored with sex pheromone trapping and RAPD (random amplified polymorphic DNA) molecular marker to analyze their movement in apple orchards. To detect their movements among farms, pheromone traps were placed at regions between apple farms (‘outside-farms’) as well as within-farms (‘inside-farms’). Four seasonal adult peaks were evident in apple-cultivating fields from April to October in both trappings of inside- or outside-farms. After overwintering generation, populations of inside-farms were significantly reduced with frequent insecticide applications, compared to populations of outside-farms. Within apple farms, G. molesta tended to be unevenly distributed because of significant sublocal preference. Active movements of local and seasonal populations of G. molesta were supported by gene flow analysis using RAPD marker. Monitoring data using sex pheromone and seasonal reduction in initial genetic differentiation detected in the overwintering populations suggest that there must be significant movement of G. molesta among different orchards in apple-cultivating areas.

벼멸구(Nilaparvata lugens)는 한국에서 월동하지 못하고 이주해 오는 종으로 서, 매년 한국의 벼 재배에 큰 손실을 주고 있다. 한국의 지역별로 채집된 벼 멸구의 유전적 변이 및 다양성을 알아보고자 마이크로새털라이트를 이용하여 실험을 수행하였다. 한국의 다섯 지역, 곤양, 광양, 진주, 남해, 남원에서 벼멸구를 각각 채집하 였고, 마이크로새털라이트 5개의 위치(18, 27, 65, 67, 76)를 기준으로 분석하 였다. 그 결과, 3개의 위치(27, 65, 67)에서 높은 유의성을 확인할 수 있었고, 한국의 벼멸구 개체 간에도 유전적으로 차이가 있음을 알 수 있었다. 마이크로새털라이트 마커를 이용한 방법은 벼멸구의 근원지 및 이동을 결 정하기 위한 실질적인 방법이 될 것으로 생각되며, 이러한 방법을 통해 비래 해충인 벼멸구의 개체군 다양성을 분석함으로서 발생 근원지와 이동경로를 파악할 수 있을 것으로 생각된다.

Geographic clines in genetic polymorphisms are widely believed as an evidence of climate change. We hypothesized green peach aphid, Myzus persicae Sulzer, one of the major insect pests in highland chinese cabbage cultivation, may also have some interactions with climate change. As the first step, we tried to find the available markers from six local strains (five collected at different heights in Hoengseong and Pyeongchang area and one from laboratory). A strain from Jeju island was used as an out-group. Although there was no significant difference in sequences of partial ribosomal RNA fragment and mitochondrial cytochrome oxidase I, and esterase isozyme pattern, we found four inter-simple sequence repeat (ISSR) markers in 22 used ISSR primers (+AGA, +CCA, +CGA, CGA+). These primers can be used as good markers to trace the M. persicae gene flow because they showed specific bands according to local strains.

베트남내 지역별로 채집된 벼멸구의 유연관계 및 개체간의 다양성을 Micro-satellite marker를 이용하여 조사하였다. 벼의 주요해충인 벼멸구(Nilaparvata lugens)는 벼를 직접 흡즙하여 피해를 줄 뿐만 아니라 2차적으로 벼에 virus병을 매개하여 벼 재배지에 심한 피해를 주고 있다. 상세한 지역 간에 유전적 변이를 확인하기 위해 베트남의 네 지역, Haipong, Ha Tay, Can Tho, Cuu long에서 채집한 벼멸구를 마이크로새털라이트 마커를 이용하여 유전적 다양성을 분석하였다. 5개의 위치(18, 27, 65, 67, 76)를 기준으로 분석한 결과, Haipong과 Ha Tay, 그리고 Can Tho와 Cuu long 지역이 각각 유전적으로 비슷할 것으로 나타났다. 이는 북쪽과 남쪽은 지리적으로 멀기 때문에 유전적 교류가 적게 일어나고, 남북의 각 지역 간에는 개체군의 이동이 보다 쉽게 발생하기 때문으로 생각된다. 이러한 정보는 베트남 지역 간에 벼멸구의 이동과 분산의 근원과 경로를 파악하는데 중요한 기초자료가 될 것으로 생각된다. 또한 비래하는 벼멸구의 근원지와 이동 경로를 분석하여 예찰함으로서 벼 재배지에 벼멸구 대발생 피해를 최소화 할 수 있을 것이다. 더 정확한 결과를 위해서는 더 많은 연구가 수행되어야 할 것으로 생각된다.

To investigate the epidemiological characteristics of 68 Listeria monocytogenes isolates, including 11 reference strains and 57 isolates from imported US beef, domestic meats (beef, pork, chicken meat), raw milk, and milk plants. L. monocytogenes was to evaluate the production of virulence proteins, such as hemolysin (LLO) and lecithinase (LCP), the adsorption of Congo red (CRA), and to detect virulence genes using the polymerase chain reaction (PCR). In the study of virulence protein production, 68(100%), 62(91.2%), and 54(79.4%) of the 68 L. monocytogenes strains were positive for LLO production, the LCP test, and the CRA test, respectively, while strains of other species, such as L. innocua, L. gray, L. murrayi, and L. welshimeri, were not. There were no significant differences between L. monocytogenes serotypes and the ability to produce LLO or LCP. L. monocytogenesstrains had very high hemolytic titers (2 to 16 fold), while the other Listeria species, other than L. ivanovii and L. seeligeri, did not. The hemolysin activities of L. monocytogenes, L. ivanovii, and L. seeligeri usually exceeded 1.0 HU/mg, while those of other Listeria spp. were less than 0.04 HU/mg. In the PCR assay, all of the L. monocytogenes strains contained the hlyA, plcA, plcB, inlA, and inlB virulence genes and produced a product of the expected size. In the PCR of the actA gene, the expected 385-bp product was seen in 39(57.4%) L. monocytogenesstrains, while an unexpected 268-bp product was seen in 29(42.6%) strains. Most L. monocytogenes strains isolated from Hanwoo beef produced the 385- bp actA gene product, while strains of imported US beef usually produced the 268-bp actA gene product. By contrast, no virulence gene products were amplified in the other Listeria spp.

To estimate the genetic characteristics and cumulative power of discrimination (CPD) existing among Hanwoo (Korean cattle) and exotic foreign population (Angus, Herford, Charolais, Holstein) we used a total of 414 genomic DNAs from five breeds population (Hanwoo, Angus, Hereford, Charolais, Holstein). Genetic characteristics indices including mean allele number among loci, unbiased heterozygosity () within locus and polymorphic information content (PIC) and unbiased average heterozygosity (H) among loci in four breeds were calculated using the generated allele frequencies by each marker. The mean allele numbers for all loci ranged between 5 and 7 while heterozygosity (H) ranged from 0.75 (HW) to 0.64 (HF) among loci and across breeds heterozygosity (H) was 0.69. The generated unbiased average heterozygosity among loci in each breed was integrated to the global formula of CPD resulting in 99.71 % within the populations. The genetic variation of HW (Hanwoo) showed highest estimates among the analyzed breeds.

Recent studies on nuclear transfer and induced pluripotent stem cells have demonstrated that differentiated somatic cells can be returned to the undifferentiated state by reversing their developmental process. These epigenetically reprogrammed somatic cells may again be differentiated into various cell types, and used for cell replacement therapies through autologous transplantation to treat many degenerative diseases. To date, however, reprogramming of somatic cells into undifferentiated cells has been extremely inefficient. Hence, reliable markers to identify the event of reprogramming would assist effective selection of reprogrammed cells. In this study, a transgene construct encoding enhanced green fluorescent protein (EGFP) under the regulation of human Oct4 promoter was developed as a reporter for the reprogramming of somatic cells. Microinjection of the transgene construct into pronuclei of fertilized mouse eggs resulted in the emission of green fluorescence, suggesting that the undifferentiated cytoplasmic environment provided by fertilized eggs induces the expression of EGFP. Next, the transgene construct was introduced into human embryonic fibroblasts, and the nuclei from these cells were transferred into enucleated porcine oocytes. Along with their in vitro development, nuclear transfer embryos emitted green fluorescence, suggesting the reprogramming of donor nuclei in nuclear transfer embryos. The results of the present study demonstrate that expression of the transgene under the regulation of human Oct4 promoter coincides with epigenetic reprogramming, and may be used as a convenient marker that non-invasively reflects reprogramming of somatic cells.

The soybean Kunitz trypsin inhibitor (KTI) protein is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. Ti locus controls presence or absence of Kunitz trypsin inhibitor protein. Genetic recombination or tight linkage between Ti locus and Satt228 marker that has been identified to be tightly linked to the Ti locus was detected for marker assisted selection (MAS) using two F2 populations of titi genotype in this study. Two F2 populations were developed from the cross of A29 (KTI protein present, TiTi genotype, AA genotype in Satt228 marker) x Gaechuck#1 and Gaechuck#2 (KTI protein absent, titi genotype, BB genotype in Satt228 marker). Among 31 F2 plants derived from A29 x Gaechuck#1, twenty nine F2 plants show BB genotype that indicates no recombination between Satt228 marker and Ti locus. Only 2 F2 plants show AA genotype that indicates recombination between Satt228 marker and Ti locus. Twenty eight F2 plants derived from A29 x Gaechuck#2 show BB genotype that indicates no recombination between Satt228 marker and Ti locus. Expected genetic ratio between Satt228 marker and Ti locus was 3.6 cM in F2 population.

한국꿩 (Korean ring-necked Pheasant, Phasianus colchicus karpowi)과 외국 아종의 유전적 유연관계를 파악하기 위해 야생 한국꿩, 사육 한국꿩, 사육 한국꿩과 외국꿩간의 잡종꿩, 외국꿩 4아종(중국 링넥, 흑 뮤탄트, 백 뮤탄트, 녹치)을 대상으로 ISSR 표지자 분석과 AMOVA 분석을 수행하였다. 야생 한국꿩의 전체 유전 다양성중 94.08%가 서식지 내 개체간 유전적 차이에 기인하고, 5.9% (Φ

Hedera helix 11계통, Hedera rhombea 3계통, Fatshedera lizei 1계통, 그리고 Fatsia japonica 1계 통을 수집하였다. RAPD primer 10개를 이용하여 수 집된 3속의 재료들의 유전적 다양성을 측정하였다. 3 속을 재료로 사용하여 밴드간 96.9%의 높은 다형성을 보였다. 총 97개의 RAPD 밴드를 이진화하여 UPGMA 방법을 이용하여 계통도를 작성하였다. Hedera helix 계통들은 모두 1개의 그룹에 속하였으며 8계통의 유전 적 거리는 극도로 적었으며 나머지 3계통 역시 유전적 으로 가까웠다. 하지만 형태적으로는 높은 다형성을 보 였다. 따라서 수집된 유전자원을 이용한 돌연변이체 개 발이 가능성이 있는 방법으로 제시되었다. Fatsia japonica는 유전적으로 관계성이 적어 다른 종들과 평 균 0.63의 유전적 거리를 보였다. Hedera helix와 Fatsia japonica 속간 교배를 통하여 개발된 Fatshedera lizei는 Hedera rhombea 계통들과 함께 계통도에서 위치하였다.

조직배양묘인 Dtps. pulcherrima (sib.) × Dtps. ‘Kyotoprinty’ 를 이용하여 호접란 체세포 변이체의 형태적 유전 적 변이를 조사하였다. 잎과 꽃에서의 색깔과 형태 변이 도 다양하게 관찰되었다. 조직배양시에 발생하는 잎의 변이와 꽃의 변이중에서 대표적인 개체들 13개를 골라 정상적인 개체와 함께 total genomic DNA를 분리하고 RAPD 분석하여 변이체와 정상적인 개체간 DNA 밴드 차이가 나는지 살펴보았다. 그 결과 Operon primer OPO-06과 OPO-07를 이용한 PCR 분석에서 다형성이 관찰되었다. 잎의 변이를 일으킨 개체와 꽃의 변이를 야 기한 개체에서 각각 특이한 DNA Marker를 선발할 수 있었다.