• 식물조직배양학회지
• /
• 제21권5호
• /
• pp.315-318
• /
• 1994

CaMV35S promoter-$\beta$-glucuronidase (GUS) 유전자와 선발 표지로서 neomycin phosphotransferase 유전자를 가진 pBI121 binary 벡터를 도입한 Agrobacterium turmfaciens LAB4404와 더덕 유식물체의 자엽 절편을 1 mg/L BA가 첨가된 MS 배지에서 48시간 동안 공동배양한 후 1 mg/L BA, 250 mg/L carbenicillin 100 mg/L kanamycin sulfate을 첨가한 고체배지에 옮겨 명조건에서 배양하였다. 배양 2주 후 절편의 절단면 부근으로부터 수많은 부정아가 형성되기 시작하였다. 이들 부정아의 GUS 활성을 조사한 결과 15%가 양성반응을 나타내었다. 배양 6주 후 절편이 형성한 수많은 부정아로부터 56.7% 빈도로 shoot이 발달하였다. 이들 shoot은 기본배지에 옮겨서 4주 경과되었을 때 대부분 발근하였으며, 재분화된 개체는 토양에 이식하였다. Southem 분석결과 GUS양성을 보인 재분화 개체의 게놈 DNA에 GUS 유전자가 삽입되었음이 확인되었다.

• 식물조직배양학회지
• /
• 제27권3호
• /
• pp.219-226
• /
• 2000

야생종 토마토의 자가불화합성에 관여하는 S RNase 유전자를 자가화합성인 재배종 토마토에 도입하여 형질전환 식물체를 만들었다. 재조합 Ti-plasmid pBI-S$_{11}$을 제작하여 Agrobacteria를 이용한 leaf disc transformation방법으로 재배종 토마토에 형질전환시켰으며, Southern분석을 통해 형질전환 식물체의 염색체 내에 자가불화합성 유전자가 안정된 구조로 도입되었는지 확인하였다. 형질전환 식물체의 화주 단백질을 이용하여 RNase활성 염색을 하여 화주 내 5단백질의 활성을 확인하였다. 또한 형질전환 식물체의 화주 단백질을 이용하여 western분석을 수행한 결과, 야생종 토마토 유래의 자가불화합성 유전자에 의해 5단백질을 정확하게 발현하는 식물체를 확인, 선발하였다. 선발된 형질전환 식물체의 화분, 화주, 화변, 잎, 줄기 등 5개의 조직을 western분석하여 5 RNase 유전자가 야생종 토마토에서와 유사한 형태로 화주조직에서만 높게 발현되며 형질전환된 재배종 토마토에서도 조직 특이적으로 발현한다는 것을 확인하였다. 위의 결과로 야생종 토마토에서 자가불화합성에 결정적인 영향을 미치는 S RNase를 재배종 토마토에 발현시켰을 때 자가불화합성을 나타내는 재배종 토마토의 표현형은 변화가 없다는 것과 어떠한 형태적인 변화도 없다는 사실을 확인하였다. 그리고 배우자체 자가불화합성 기작에는 화주쪽의 S RNase 이외에 화분쪽의 또 다른 기구가 있다는 것을 추측할 수 있었다.

• 한국정보통신학회논문지
• /
• 제11권7호
• /
• pp.1318-1324
• /
• 2007

지금까지의 상품 검색 방법으로는 찾고자하는 정보를 검색할 때 주로 단어의 빈도수나 어휘 정보를 이용하는 키워드 기반의 검색이 주로 쓰이고 있었다. 키워드 기반의 검색에서는 사용자의 질의와 관련이 없는 문서들까지도 같은 결과로 나타내 주고 이로 인해 사용자는 제시된 결과를 한번 더 수동적으로 검색해야하는 부담을 않게 되었다. 이러한 문제점을 해결하기 위해 온톨로지가 대두되었다. 본 논문에서는 온톨로지를 이용한 상품 검색 시스템을 직접 구축하여 분류별 검색을 통해 얼마나 정확한 검색을 하는지 실험하였다. 실험을 위해 전국적으로 On/Off라인 할인점을 운영 중에 있는 A할인점의 상품 데이터 약 40,000여개를 데이터 베이스로 구축하였고 User Interface 개발환경은 JSP와 PowerBuilder9.0을 사용하여 검색 시스템을 개발하여 실험하였다. 그 결과 본 논문에서 제안하고 설계한 상품 도메인 온톨로지를 이용한 검색 방법이 기존의 기반의 검색 방법보다 우수한 결과를 나타내고 있음을 입증하였다.

• Mycobiology
• /
• 제47권3호
• /
• pp.301-307
• /
• 2019

The $11{\alpha}$-hydroxylation of $16{\alpha}$, 17-epoxyprogesterone (EP) catalyzed by Rhizopus nigricans is crucial for the steroid industry. However, lower conversion rate of the biohydroxylation restricts its potential industrial application. The $11{\alpha}$-steroid hydroxylase CYP509C12 from R. oryzae were reported to play a crucial role in the $11{\alpha}$-hydroxylation in recombinant fission yeast. In the present study, the CYP509C12 of R. oryzae (RoCYP) was introduced into R. nigricans using the liposome-mediated mycelial transformation. Heterologous expression of RoCYP resulted in increased fungal growth and improved intracellular reactive oxygen species content in R. nigricans. The $H_2O_2$ levels in RoCYP transformants were approximately 2-folder that of the R. nigricans wild type (RnWT) strain, with the superoxide dismutase activities increased approximately 45% and catalase activities decreased approximately 68%. Furthermore, the $11{\alpha}$-hydroxylation rates of EP in RoCYP transformants (C4, C6 and C9) were 39.7%, 38.3% and 38.7%, which were 12.1%, 8.2% and 9.4% higher than the rate of the RnWT strain, respectively. This paper investigated the effect of heterologous expression of RoCYP in R. nigricans, providing an effective genetic method to construct the engineered strains for steroid industry.

• 대한임상검사과학회지
• /
• 제52권4호
• /
• pp.310-316
• /
• 2020

마이크로어레이 진단 기법의 발달은 세포유전학적 관점에서, 다양한 종류의 유전학적 질병과 관련하여 새로운 정보를 제공하고, 질병에 대한 기본적인 통찰력을 제공하는데 매우 중요한 역할을 제공하고 있다. 그동안 많은 연구들에서, 마이크로어레이 기술을 활용한 인간 게놈의 유동성과 다양성을 입증해 주었으며, 게놈의 취약성을 식별하기 위한 보다 정확한 진단기법과 적절한 임상 관리 방법을 효율적으로 제공해 왔다. 앞으로 다양한 유전과 관련된 질병에 기존 세포유전학적 방법을 자동화된 마이크로어레이 방법으로 전환한다면, 보다 효율적인 방법으로 질병을 진단하고, 정확성을 향상시키며, 유전자 배열의 암호화 및 복잡한 특성을 밝히는데 매우 중요한 역할을 할 것으로 생각된다. 또한 이 분석 기법을 활용하여 게놈과 인간의 건강, 질병과의 관계를 분석하여 다양한 정보를 미리 제공하여 질병을 예방하고, 질병의 진단 및 치료에도 도움이 될 수 있는 새로운 혁명을 일으킬 수 있을 것으로 기대된다.

• Journal of Plant Biotechnology
• /
• 제49권2호
• /
• pp.131-138
• /
• 2022

An efficient genetic transformation protocol is a fundamental requirement for high regeneration capacity from cultivated durum wheat (Triticum durum) varieties. In this study, wereportedtheeffectsoftwoauxins,2,4-dichlorophenoxyaceticacid(2,4-D)and4-amino-3,5,6-trichloropicoli nicacid(picloram), at a concentration of 2 mg/Laloneandincombination on the embryogenic callus and plantlet regeneration of four durum wheat varieties (Amria, Chaoui, Marouane, and Tomouh) using mature embryos (MEs) and immature embryos (ImEs). Significanteffectsofvariety,culturemedium(theauxinused),andvariety-mediuminteraction were observed on the callus weight and plantlet regeneration of both MR and ImE explants. The medium used for callus induction significantly affected plantlet regeneration (p < 0.001). Comparedto2,4-D, picloram led to a higher plantlet regeneration rate in both ME and ImE explants (19.8% and 40.86%, respectively). Plantlet regeneration also varied significantly depending on the variety and medium used. PicloramledtohighplantletregenerationofbothME and ImE explants in all varieties except Tomouh, which showed high plantlet regeneration of ME explants in 2,4-D. A comparison of ME and ImE responses indicated that ImEs are the best explants for high plantlet regeneration in durum wheat. Ourfindingssuggestthatpicloramisthebestauxin and should be used instead of 2,4-D due to its positive effect on increasing plant regeneration of durum wheat ME and ImE explants.

• Journal of Yeungnam Medical Science
• /
• 제40권3호
• /
• pp.283-288
• /
• 2023

Severe chronic neutropenia is classified as severe congenital, cyclic, autoimmune, or idiopathic. However, there is a lot of uncertainty regarding the diagnosis of severe congenital neutropenia (SCN) and chronic idiopathic neutropenia, and this uncertainty affects further evaluations and treatments. A 20-year-old man presented with fever and knee abrasions after a bicycle accident. On admission, his initial absolute neutrophil count (ANC) was 30/µL. He had no medical history of persistent severe neutropenia with periodic oscillation of ANC. Although his fever resolved after appropriate antibiotic therapy, ANC remained at 80/µL. Bone marrow (BM) aspiration and biopsy were performed, and a BM smear showed myeloid maturation arrest. Moreover, genetic mutation test results showed a heterozygous missense variant in exon 4 of the neutrophil elastase ELANE: c597+1G>C (pV190-F199del). The patient was diagnosed with SCN. After discharge, we routinely checked his ANC level and monitored any signs of infection with minimum use of granulocyte colony-stimulating factor (G-CSF), considering its potential risk of leukemic transformation. Considering that SCN can be fatal, timely diagnosis and appropriate management with G-CSF are essential. We report the case of a patient with SCN caused by ELANE mutation who had atypical clinical manifestations. For a more accurate diagnosis and treatment of severe chronic neutropenia, further studies are needed to elucidate the various clinical features of ELANE.

• Journal of Animal Science and Technology
• /
• 제66권2호
• /
• pp.279-294
• /
• 2024

Agriculture has played a significant role in the national economy, contributing to food security, driving economic growth, and safeguarding the dietary habits of the population. Korean agriculture has been compelled to focus on intensive farming due to its limited cultivation area, excessive input costs, and the limitations of agricultural mechanization. In the Republic of Korea (R.O.K), the concept of environmentally friendly animal agriculture began to be introduced in the early 2000s. This concept ultimately aims to cultivate sustainable animal agriculture (SAA) through environmentally friendly production practices, ensuring the healthy rearing of animals to supply safe animal products. Despite the government's efforts, there are still significant challenges in implementing environmentally friendly agriculture and SAA in the R.O.K. Therefore, the objective of this review is to establish the direction that the animal agriculture sector should take in the era of climate crisis, and to develop effective strategies for SAA tailored to the current situation in the R.O.K by examining the trends in SAA in the U.S. The animal agriculture sector in the U.S. has been working towards creating a SAA system where humans, animals, and the environment can coexist through government initiatives, industry research, technological support, and individual efforts. Efforts have been made to reduce emissions like carbon, and improve factors affecting the environment such as the carbon footprint, odor, and greenhouse gases associated with animal agriculture processes for animals such as cattle and pigs. The transition of the U.S. towards SAA appears to be driven by both external goals related to addressing climate change and the primary objectives of responding to the demand for safe animal products, expanding consumption, and securing competitiveness in overseas export markets. The demand for animal welfare, organic animal products, and processed goods has been increasing in the U.S. consumer market. A major factor in the transformation of the U.S. animal agriculture sector in terms of livestock specifications is attributed to environmentally friendly practices such as high-quality feed, heat stress reduction, improvements in reproductive ability and growth period reduction, and efforts in animal genetic enhancement.

• 한국균학회소식:학술대회논문집
• /
• 한국균학회 2015년도 춘계학술대회 및 임시총회
• /
• pp.53-53
• /
• 2015

Agrobacterium tumefaciens-mediated transformation(ATMT) of Flammulina velutipes was used to produce a diverse number of transformants to discover the functions of gene that is vital for its variation color, spore pattern and cellulolytic activity. Futhermore, the transformant pool will be used as a good genetic resource for studying gene functions. Agrobacterium-mediated transformation was conducted in order to generate intentional mutants of F. velutipes strain KACC42777. Then Agrobacterium tumefaciens AGL-1 harboring pBGgHg was transformed into F. velutipes. This method is use to determine the functional gene of F. velutipes. Inverse PCR was used to insert T-DNA into the tagged chromosomal DNA segments and conducting sequence analysis of the F. velutipes. But this experiment had trouble in diverse morphological mutants because of dikaryotic nature of mushroom. It needed to make monokaryotic fruiting varients which introduced genes of compatible mating types. In this study, next generation sequencing data was generated from 28 strains of Flammulina velutipes with different phenotypes using Illumina Hiseq platform. Filtered short reads were initially aligned to the reference genome (KACC42780) to construct a SNP matrix. And then we built a phylogenetic tree based on the validated SNPs. The inferred tree represented that white- and brown- fruitbody forming strains were generally separated although three brown strains, 4103, 4028, and 4195, were grouped with white ones. This topological relationship was consistently reappeared even when we used randomly selected SNPs. Group I containing 4062, 4148, and 4195 strains and group II containing 4188, 4190, and 4194 strains formed early-divergent lineages with robust nodal supports, suggesting that they are independent groups from the members in main clades. To elucidate the distinction between white-fruitbody forming strains isolated from Korea and Japan, phylogenetic analysis was performed using their SNP data with group I members as outgroup. However, no significant genetic variation was noticed in this study. A total of 28 strains of Flammulina velutipes were analyzed to identify the genomic regions responsible for producing white-fruiting body. NGS data was yielded by using Illumina Hiseq platform. Short reads were filtered by quality score and read length were mapped on the reference genome (KACC42780). Between the white- and brown fruitbody forming strains. There is a high possibility that SNPs can be detected among the white strains as homozygous because white phenotype is recessive in F. velutipes. Thus, we constructed SNP matrix within 8 white strains. SNPs discovered between mono3 and mono19, the parental monokaryotic strains of 4210 strain (white), were excluded from the candidate. If the genotypes of SNPs detected between white and brown strains were identical with those in mono3 and mono19 strains, they were included in candidate as a priority. As a result, if more than 5 candidates SNPs were localized in single gene, we regarded as they are possibly related to the white color. In F. velutipes genome, chr01, chr04, chr07,chr11 regions were identified to be associated with white fruitbody forming. White and Brown Fruitbody strains can be used as an identification marker for F. veluipes. We can develop some molecular markers to identify colored strains and discriminate national white varieties against Japanese ones.

• 식물병연구
• /
• 제19권4호
• /
• pp.254-258
• /
• 2013

후자린산(FA)는 Fusarium 속 균이 생성하는 독소로서 다른 곰팡이독소보다 독성은 낮으나 다른 독소와 중복 오염시 전체 독성을 증진시키는 것으로 알려졌다. 현재까지 FA 생합성 관련 효소나 유전자가 Fusarium oxysporum에서 밝혀지지 않았기 때문에 본 연구에서는 관련 생합성 유전자의 발굴을 위해 제한효소를 통한 무작위 삽입 형질전환방법인 REMI를 이용하여 FA 생성 F. oxysporum 균주의 생합성유전자의 결손을 시도하였다. F. oxysporum 균주 2주를 대상으로 REMI를 시도한 결과, 평균 3.2주 ($1{\mu}g$ DNA 당)의 효율로 7,100주 이상의 형질전환체를 육성하였다. FA 미생성 형질전환체를 스크리닝 하기 위해 FA가 함유된 배양액에서 다양한 식물종자의 발아여부를 조사한 결과, 11종의 종자 중 가장 감수성인 배추종자를 선발하였다. 각 형질전환체는 Czapek-Dox broth에서 3주간 배양한 후 배양여액을 배추종자의 발아여부 검정에 사용하였다. 검정결과 총 5,000여 주의 REMI 형질전환체 중 53주의 배양여액에서 종자가 발아하지 않아, 이들을 FA 미(저) 생성 추정 형질전환체로 선발하였다. 이중 26주의 FA 생성량을 HPLC로 분석한 결과, 2주의 형질전환체에서 모균주 생성량의 1% 이하의 FA가 검출되었다. 이 중 형질전환체 1주로부터 REMI 벡터 삽입 부위 게놈 DNA의 염기서열(252 bp)을 확보하였으며, 이 부위는 F. fujikuroi의 미동정 게놈부위와 93% 유사성이 있음을 확인하였다. 이 부위의 FA생성 관련성 증명을 위해서는 추후 연구가 필요하다.