• BMB Reports
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• 제34권6호
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• pp.502-508
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• 2001

A cDNA, encoding the human growth hormone (hGH), was synthesized based on the known 191 amino acid sequence. Its codon usage was optimized for a high level expression in Escherichia coli. Unique restriction sites were incorporated throughout the gene to facilitate mutagenesis in further studies. To minimize an initiation translation problem, a 624-bp cassette that contained a ribosome binding site and a start codon were fused to the hGH-coding sequence that was flanked between the EcoRI and HindIII sites. The whole fragment was synthesized by an overlapped extension of eight long synthetic oligonucleotides. The four-short duplexes of DNA, which were first formed by annealing and filling-in with a Klenow fragment, were assembled to form a complete hGH gene. The hGH was cloned and expressed successfully using a pET17b plasmid that contained the T7 promoter. Recombinant hGH yielded as much as 20% of the total cellular proteins. However, the majority of the protein was in the form of insoluble inclusion bodies. N-terminal amino acid sequencing also showed that the hGH produced in E. coli contained formyl-methionine. This study provides a useful model for synthesis of the gene of interest and production of recombinant proteins in E. coli.

• Journal of Microbiology and Biotechnology
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• 제20권3호
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• pp.630-636
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• 2010

A survey of the endangered orchid Orchis militaris populations was carried out in north-eastern Italy. The occurrence of fungal root endophytes was investigated by light and electron microscopies and molecular techniques. Two main sites of presence were individuated in the Euganean Hills, differing as to the percentage of flowering individuals and of capsules completing maturity. Fluorescence microscopy revealed an intracellular cortical colonization by hyphal pelotons. Two ITS PCR products co-amplified. Sequencing revealed for the former an identity and a high similarity (99%) with a Tulasnellaceae (Basidiomycota) fungus found within tissues of the same host in independent studies in Hungary and Estonia, suggesting an interesting case of tight specificity throughout the Eurosiberian home range. The second amplicon had 99% similarity with Tetracladium species (Ascomycota) recently demonstrated as potential endophytes. TEM revealed two different hyphal structures. Double fungal colonization appears to occur in Orchis militaris and the possible requirement of a specific fungal partner throws light on the causes of this plant's rarity and threatened status.

• Fisheries and Aquatic Sciences
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• 제24권9호
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• pp.297-310
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• 2021

The rockworm gut flora was identified at each growth stage to elucidate the composition of the bacterial community. The source material was Marphysa sanguinea fed regular feed and fed biofloc at the adult stage in parallel. The systematic bacterial community composition was determined based on the next-generation sequencing method, and alpha diversity and beta diversity were conducted to access the species diversity within and between the bacterial communities, respectively. The composition of the gut flora changed considerably as the rockworms developed. The shift in the gut flora was confirmed at the phylum, family, and genera level of the bacterial communities. The Vibrio species associated with high rockworm mortality occupied 7.7% of the gut flora at the larval stage; however, they disappeared in the healthy adult gut. Moreover, different gut flora was observed between adults fed regular feed and those fed biofloc. Specifically in the biofloc-fed adult gut, several immune relevant and water-purifying bacteria were detected. The biofloc-fed adult gut flora could decompose and mineralize organic sediment, and thus be effectively utilized for integrated multitrophic aquaculture. The Venn diagram revealed that only two bacterial species were shared throughout all growth stages, and the biofloc-fed adults exhibited the highest diversity within the bacterial community.

• 식물병연구
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• 제23권3호
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• pp.262-267
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• 2017

2015년부터 2016년까지 국내 5개 지역에서 기장 101점, 수수 200점의 시료를 채집하였다. 채집한 시료에 대해 paired-end RNA sequencing, RT-PCR 진단을 수행하였다. 그 결과, 경상도에서 채집한 수수에서 Rice black-streaked dwarf virus (RBSDV)가 검출되었으며, RBSDV, Rice stripe virus (RSV), Barley virus G (BVG), Cereal yellow dwarf virus (CYDV) 4종의 바이러스가 기장에서 검출되었다. 기장에서 검출된 4종의 바이러스 중 RSV와 RBSDV는 경상도에서 채집한 시료에서 높은 감염률을 보였다. 반면, BVG는 5개 지역에서 모두 감염이 확인되었으며, 이미 전국적으로 분포되어 있는 바이러스인 것으로 보인다. 본 연구에서는 국내 기장과 수수에서 RBSDV를 처음으로 동정하였으며, 수수에서 검출된 CYDV는 기존에 보고된 CYDV 분리주와 상동성이 상대적으로 낮은 것으로 보아 Polerovirus 속의 신종 바이러스로 예상된다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권3호
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• pp.320-333
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• 2019

Objective: The Shanghai Holstein cattle breed is susceptible to severe mastitis and other diseases due to the hot weather and long-term humidity in Shanghai, which is the main distribution centre for providing Holstein semen to various farms throughout China. Our objective was to determine the genetic mechanisms influencing economically important traits, especially diseases that have huge impact on the yield and quality of milk as well as reproduction. Methods: In our study, we detected the structural variations of 1,092 Shanghai Holstein cows by using next-generation sequencing. We used the DELLY software to identify deletions and insertions, cn.MOPS to identify copy-number variants (CNVs). Furthermore, we annotated these structural variations using different bioinformatics tools, such as gene ontology, cattle quantitative trait locus (QTL) database and ingenuity pathway analysis (IPA). Results: The average number of high-quality reads was 3,046,279. After filtering, a total of 16,831 deletions, 12,735 insertions and 490 CNVs were identified. The annotation results showed that these mapped genes were significantly enriched for specific biological functions, such as disease and reproduction. In addition, the enrichment results based on the cattle QTL database showed that the number of variants related to milk and reproduction was higher than the number of variants related to other traits. IPA core analysis found that the structural variations were related to reproduction, lipid metabolism, and inflammation. According to the functional analysis, structural variations were important factors affecting the variation of different traits in Shanghai Holstein cattle. Our results provide meaningful information about structural variations, which may be useful in future assessments of the associations between variations and important phenotypes in Shanghai Holstein cattle. Conclusion: Structural variations identified in this study were extremely different from those of previous studies. Many structural variations were found to be associated with mastitis and reproductive system diseases; these results are in accordance with the characteristics of the environment that Shanghai Holstein cattle experience.

• 한국자원식물학회지
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• 제36권4호
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• pp.290-298
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• 2023

본 연구에서는 닥나무 속 식물에 대한 InDel 마커를 개발하였다. 전국의 닥나무 속 식물 22개체를 수집하였고, 수집한 닥나무 속 식물 중 6개체를 차세대염기서열 분석(NGS)을 실시하였다. NGS를 통하여 얻은 염기서열 정보를 기존에 발표되었던 닥나무 엽록체 서열과 비교하여 InDel 마커 후보를 선발하였다. 선발한 마커 후보를 수집된 닥나무 속 식물에 적용하여 마커의 특성 검정을 통해 총5개의 엽록체 기반 마커를 개발하였다. 개발된 InDel 마커를 22개의 유전자원에 적용한 후 군집 분석을 실시한 결과, 총5개의 그룹으로 나뉘었다. 본 연구에서 개발된 마커들은 닥나무 속의 육종이나 종 판별에 활용할 수 있을 것이라 판단된다.

• Journal of Ginseng Research
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• 제44권1호
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• pp.123-134
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• 2020

Background: The lepidopteran Asiatic corn borer (ACB), Ostrinia furnacalis (Guenee), has caused huge economic losses throughout the Asian-Western Pacific region. Usually, chemical pesticides are used for the control, but excessive use of pesticides has caused great harm. Therefore, the inartificial ecotypic pesticides to ACB are extremely essential. In our previous study, we found that panaxadiol saponins (PDS) can effectively reduce the harm of ACB by causing antifeedant activity. Therefore, it is necessary to reveal the biological molecular changes in ACB and the functionary mechanism of PDS. Methods: We analyzed the global transcription of ACB with different PDS concentration treatment (5 mg/mL, 10 mg/mL, and 25 mg/mL) by high-throughput sequencing and de novo transcriptome assembly method. Results: PDS treatment could cause the changes of many gene expressions which regulate its signal pathways. The genes in peroxisome proliferator-activated receptor (PPAR) signaling pathway were significantly downregulated, and then, the downstream fatty acid degradation pathway had also been greatly affected. Conclusion: Through this experiment, we hypothesized that the occurrence of antifeedant action of ACB is because the PDS brought about the downregulation of FATP and FABP, the key regulators in the PPAR, and the downregulation of FATP and FABP exerts further effects on the expression of SCD-1, ACBP, LPL, SCP-X, and ACO, which leads to the disorder of PPAR signaling pathway and the fatty acid degradation pathway. Not only that, PDS treatment leads to enzyme activity decrease by inhibiting the expression of genes associated with catalytic activity, such as cytochrome P450 and other similar genes.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.441-453
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• 2019

Organic farming is considered an effective form of sustainable agricultural management. However, understanding of soil microbial diversity and composition under long-term organic and conventional farming is still limited and controversial. In this study, the Illumina MiSeq platform was applied to investigate the responses of soil bacterial and fungal diversity and compositions to organic farming (OF) and improved conventional farming (CF, applied straw retention) in the rice-wheat rotation system. The results highlighted that the alpha diversity of microbial communities did not differ significantly, except for higher bacterial diversity under OF. However, there were significant differences in the compositions of the soil bacterial and fungal communities between organic and conventional farming. Under our experimental conditions, through the ecological functional analysis of significant different or unique bacterial and fungal taxonomic members at the phyla and genus level, OF enhanced nitrogen, sulfur, phosphorus and carbon dynamic cycling in soil with the presence of Nodosilinea, Nitrospira, LCP-6, HB118, Lyngbya, GOUTA19, Mesorhizobium, Sandaracinobacter, Syntrophobacter and Sphingosinicella, and has the potential to strengthen soil metabolic ability with Novosphingobium. On the other hand, CF increased the intensity of nitrogen cycling with Ardenscatena, KD1-23, Iamia, Nitrosovibrio and Devosia, but enriched several pathogen fungal members, including Coniochaeta, Corallomycetella, Cyclaneusma, Cystostereum, Fistulina, Curvularia and Dissoconium.

• 미생물학회지
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• 제53권1호
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• pp.1-8
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• 2017

홍역바이러스(Measles virus)는 전염성이 매우 높은 발진성 열병을 일으키는 바이러스이며, 약독화된 생백신 접종으로 충분히 예방할 수 있다. 그러나 대부분의 개발도상국에서 어린아이들의 주요 사망원인이며 해마다 20만명이 사망하는 풍토병이다. 우리나라는 1996년 이후부터 국가적으로 시행된 정기예방접종으로 인한 높은 백신 접종률(>95.0%)로 획득된 집단면역과 고도의 기술적 감시체계의 결과로 홍역이 퇴치되었으며, 2014년 3월 WHO로부터 '홍역퇴치국가'로 인증 받았다. 그러나 2014년 초부터 경기도에서 홍역환자가 다시 발생하였다. 본 연구에서는 2014년 1월부터 2014년 7월까지 경기도에서 발생한 홍역 확진 환자를 대상으로 홍역바이러스의 유행양상 및 유전형 분포양상을 분석하였다. 72건의 홍역바이러스 양성검체에서 60주의 홍역바이러스를 분리하였고, nucleoprotein(N) 유전자의 염기서열 분석을 통하여 유전형의 확인 및 유전자변이를 분석하였다. 총 60건의 홍역 확진검체를 분석한 결과 58건(96.7%)에서 B3 genotype을 확인하였다. 또한 nucleoprotein(N) 유전자 부위의 유전자 변이 분석결과, 경기도내 분리주들이 2013년에서 2014년 필리핀에서 유행한 홍역 분리주들과 99% 일치하였다. 따라서 필리핀 등에서 감염된 후 국내에 유입되어 2차 감염이 된 것으로 사료된다.

• 생명과학회지
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• 제28권9호
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• pp.1118-1126
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• 2018

본 논문은 양식 어류에게 많은 경제적 피해를 입히고 있는 다양한 어류질병세균 중에서 연쇄구균증에 대한 일반적인 특성과 종류, 진단 방법에 대하여 기존에 연구 보고된 논문을 기반으로 알아보고자 한다. 대표적인 어류연쇄구균증의 원인체로는 Lactococcus garvieae, Streptococcus iniae, S. parauberis가 있다. 연쇄구균증에 감염 시 나타나는 증상으로는 체색변화, 안구이상, 아가미 퇴색, 출혈, 복부팽만, 신장과 비장의 종대 등의 보이다 폐사가 일어난다. 또한 수온이 상승하는 6월부터 10월까지 주로 일어나며 집단적으로 발병하여 폐사가 일어난다. 현재 연쇄구균증을 진단하기 위한 기술로는 16S rRNA, 16S-23S rRNA intergenic spacer region (ISR), Random Amplified polymorphic DNA (RAPD), Ribotyion (RT), Loop-mediated isothermal amplification (LAMP) 등이 있다. 이중에서 현장에서 적용 가능성이 높은 LAMP법이 각광을 받고 있지만 결과 확인 등의 문제로 인하여 현재는 어려움을 겪고 있는 실정이다. 이에 현장에서 진단부터 결과 확인까지 손쉽게 사용할 수 있도록 문제점을 보완하면 연쇄구균증에 대한 경제적 손실을 최소화 할 것으로 사료된다.