• Title/Summary/Keyword: artificial gene

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Lower Antibody Response in Chickens Homozygous for the Mx Resistant Allele to Avian Influenza

  • Qu, L.J.;Li, X.Y.;Xu, G.Y.;Ning, Z.H.;Yang, N.
    • Asian-Australasian Journal of Animal Sciences
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    • 제22권4호
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    • pp.465-470
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    • 2009
  • The chicken Mx gene has been regarded as a candidate gene for resistance to avian influenza virus (AIV). In this study, three groups of chickens with homozygotes (AA, GG) and heterozygotes (AG) of the resistant (A) and susceptible alleles (G) to AIV of the Mx gene were constructed from a line of dwarf egg-type chickens. These chickens were not examined for their resistant activities to AIV because the differential resistance had only been detected in vitro. The birds of the three groups were vaccinated with inactivated H5N2 AIV vaccine and the level of hemagglutination inhibition (HI) antibody to AIV was detected. The association between disease resistant activity to AIV and antibody response to AIV vaccination in the three groups was analyzed. The chickens with homozygous resistant allele A showed the lowest antibody levels, whereas the heterozygous chickens (AG) presented the highest antibody level after the boosting vaccination, which indicates that the efficiency of artificial selection on the resistant allele of Mx gene will be compromised since the homozygotes of the allele presented the weakest antibody response to the corresponding vaccine.

합성 유전자를 이용하여 Escherichia coli에서 백신 후보의 생산 혹은 진단용 항체의 개발을 위한 인간 rotavirus VP8* 부분 단백질의 발현 (Use of the Synthetic Gene Encoding the Truncated Human Rotavirus VP8* Protein in Escherichia coli for Production of Vaccine Candidates or Development of Diagnostic Antibodies)

  • 김상래;이병욱
    • 생명과학회지
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    • 제28권4호
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    • pp.478-482
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    • 2018
  • 인간 rotavirus는 영아에게 급성 설사를 일으키는 병원체의 하나이다. 본 연구에서는 Escherichia coli의 코돈 선호도를 따라서 인간 rotavirus A (serotype 1 strain WA)의 $VP8^*$ 단백질을 일부분 암호화하도록 인공적인 유전자를 합성하였다. 합성된 $VP8^*$ 유전자는 코돈을 번역틀에 일치시키고 클로닝이 용이하도록 하기 위한 NdeI 및 HindIII 제한효소 절단 부위와 친화적 정제를 위한 6-히스티딘 암호화 서열을 C-말단에 보유하고 있다. 합성된 $VP8^*$ DNA 절편을 pT7-7 발현 벡터에 삽입하여 E. coli BL21 (DE3)로 형질전환한 후에 최종 농도 0.05 mM IPTG로 생산을 유도한 결과 예상했던 대로 19.7-kDa 크기의 $VP8^*$ 단백질이 고농도로 발현되었다. SDS-PAGE에 전개된 단백질들을 대상으로 mouse anti-rotavirus capsid antibody를 사용한 Western blotting의 결과 ~20-kDa $VP8^*$ 단백질 밴드가 관찰되었다. 인공 $Vp8^*$ 단백질이 피하 주사된 토끼의 polyclonal antibody 혈장을 이용한 조사에서도 동일한 크기의 단백질 밴드를 확인할 수 있었다. 이는 합성된 유전자가 바이러스성 질환을 통제할 항원성 백신 후보의 생산 혹은 진단용 항체를 개발하기 위한 쉽고 빠른 방법을 제공할 수 있다는 의미이다.

Novel Alternative Methods in Toxicity Testing

  • Satoh, Tetsuo
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1994년도 춘계학술대회 and 제3회 신약개발 연구발표회
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    • pp.129-130
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    • 1994
  • The science of toxicology is the understanding of the mechanisms by which exogenous agents produce deleterious effects in biological systems. The actions of chemicals such as drugs are ultimately exerted at the cellular and gene levels. Over the past decade. several in vitro alternative methods such as cultured cells for assessing the toxicity of various xenobiotics have been proposed to reduce the use of animals. In this workshop three advanced methods will be presented. These methods are novel important models for toxicologic studies. Dr. Tabuchis group has establishcd two immortalized gastric surface mucosa cell lines from the pminary cultore of gastric fundic mucosal cells of adult transgenic mice harboring a temperature sensitive simian virus 40 large T-anugen gene. As the immortalized cell lines of various tissues possess unique characteristics to maintain their normal functions for several months, these cell lines are extremely useful for not only toxicity testing but also pharmacological screening in new drug development. Professor Funatsu have studied the formation of spherical multicelluar aggregates of adult rat hepatocytes(spheroid) having tissue like structure. The sphcroid shown thre is a prototype module of an artificial liver support system. Thus, the urea synthesis activity of the artificial liver was maintained at least to days in 100% rat blood plasma. Dr. Takezawa and his coworkers have developed a novel culture system of multicellular spheroids considered 〃organoids〃 by utilizing a thermo-responsive polymer as a substratum of anchorage dependent cells. His final goal is to reconstitute the organoids of various normal organs, e.g., liver, skin etc. and also abnormal deseased organs such as tumor.

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Flexural capacity estimation of FRP reinforced T-shaped concrete beams via soft computing techniques

  • Danial Rezazadeh Eidgahee;Atefeh Soleymani;Hamed Hasani;Denise-Penelope N. Kontoni;Hashem Jahangir
    • Computers and Concrete
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    • 제32권1호
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    • pp.1-13
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    • 2023
  • This paper discusses a framework for predicting the flexural strength of prestressed and non-prestressed FRP reinforced T-shaped concrete beams using soft computing techniques. An analysis of 83 tests performed on T-beams of varying widths has been conducted for this purpose with different widths of compressive face, beam depth, compressive strength of concrete, area of prestressed and non-prestressed FRP bars, elasticity modulus of prestressed and non-prestressed FRP bars, and the ultimate tensile strength of prestressed and non-prestressed FRP bars. By analyzing the data using two soft computing techniques, named artificial neural networks (ANN) and gene expression programming (GEP), the fundamental parameters affecting the flexural performance of prestressed and non-prestressed FRP reinforced T-shaped beams were identified. The results showed that although the proposed ANN model outperformed the GEP model with higher values of R and lower error values, the closed-form equation of the GEP model can provide a simple way to predict the effect of input parameters on flexural strength as the output. The sensitivity analysis results revealed the most influential input parameters in ANN and GEP models are respectively the beam depth and elasticity modulus of FRP bars.

Improving classification of low-resource COVID-19 literature by using Named Entity Recognition

  • Lithgow-Serrano, Oscar;Cornelius, Joseph;Kanjirangat, Vani;Mendez-Cruz, Carlos-Francisco;Rinaldi, Fabio
    • Genomics & Informatics
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    • 제19권3호
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    • pp.22.1-22.5
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    • 2021
  • Automatic document classification for highly interrelated classes is a demanding task that becomes more challenging when there is little labeled data for training. Such is the case of the coronavirus disease 2019 (COVID-19) clinical repository-a repository of classified and translated academic articles related to COVID-19 and relevant to the clinical practice-where a 3-way classification scheme is being applied to COVID-19 literature. During the 7th Biomedical Linked Annotation Hackathon (BLAH7) hackathon, we performed experiments to explore the use of named-entity-recognition (NER) to improve the classification. We processed the literature with OntoGene's Biomedical Entity Recogniser (OGER) and used the resulting identified Named Entities (NE) and their links to major biological databases as extra input features for the classifier. We compared the results with a baseline model without the OGER extracted features. In these proof-of-concept experiments, we observed a clear gain on COVID-19 literature classification. In particular, NE's origin was useful to classify document types and NE's type for clinical specialties. Due to the limitations of the small dataset, we can only conclude that our results suggests that NER would benefit this classification task. In order to accurately estimate this benefit, further experiments with a larger dataset would be needed.

Characterization of Vaginal Microbiota Associated with Pregnancy Outcomes of Artificial Insemination in Dairy Cows

  • Chen, Shi-Yi;Deng, Feilong;Zhang, Ming;Jia, Xianbo;Lai, Song-Jia
    • Journal of Microbiology and Biotechnology
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    • 제30권6호
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    • pp.804-810
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    • 2020
  • The profitability of the dairy and beef industries is largely affected by the actually achieved reproductive efficiency. Although a large proportion of cows worldwide are bred by artificial insemination (AI) services, many potential factors affecting the outcome of pregnancy by AI remain to be addressed. In the present study, we investigated the vaginal microbiota by high-throughput sequencing of 16S rRNA gene and analyzed their association with differential pregnancy outcomes (i.e., pregnant vs. nonpregnant) of multiple AI services in dairy cows. Sequencing of the V3-V4 region totally produced 512,046 high-quality sequences that were computationally clustered into 2,584 operational taxonomic units (OTUs). All OTUs were taxonomically assigned to 10 bacterial phyla. There were statistically significant differences among the three AI service times (T1, T2 and T3) with respect to the Shannon index and number of observed OTUs (p < 0.05). Bray-Curtis distance-based PCoA analysis also revealed that T2 group could be significantly distinguished from T1 and T3. However, no significant difference between the pregnant and nonpregnant cows was found in confidence regarding both alpha diversity and beta diversity. These results could help us better understand the possible influence of vaginal microbial community on pregnancy outcomes of AI service in cows.

초파리의 보행행동에 관한 인위도태와 자연도태에 의한 유전적 효과 (Effects of Artificial and Natural Selection on Walking Behavior in Drosophila melanogaster)

  • 주종길;이현화
    • 한국동물학회지
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    • 제26권2호
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    • pp.95-106
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    • 1983
  • Drosophila melanogaster의 Oregon-R 계통과 lethal free 집단을 대상으로 connected test tube apparatus를 사용하여 보행행동에 관한 rapid와 slow 행동을 방향성도태의 방법으로 15세대 동안에 걸쳐 도태하였다. 한편 10세대째부터 natural selection을 행하여 유전적 효과를 분석하였다. 1. 보행행동의 rapid와 slow 성질은 초기세대에서부터 뚜렷한 도태효과를 나타내어 제 7세대 이후에 각각 selection plateau에 달하였다. 2. 방향성 도태를 10세대 동안 실시한 후 realized heritability를 계산한 결과 rapid 성질은 $9\\sim14%$, slow 성질은 $11\\sim16%$로서 rapid행동보다 slow 행동의 유전율이 다소 높게 나타났다. 3. Rapid 성질을 지배하는 유전자와 slow 성질을 지배하는 유전자의 우열관계를 밝히기 위한 hybridization 실험결과 slow 유전자가 rapid 유전자에 대하여 partial dominance의 효과가 있었다. 4. 10세대 동안에 걸쳐 방향성 도태를 실시한 후 natural selection을 5세대 동안 실시한 결과 rapid 성질은 단 5세대만에 neutral의 상태 (6.5)로 복원되었으나 slow 성질은 모집단의 보행지수와 비교하여 전혀 변화가 없었다. 실험결과로 미루어 rapid와 slow 형질은 polygenic system에 의하여 control 되는 양적 형질임을 알았다. 한편 rapid 유전자는 natural selection에 의한 homeostasis의 효과가 있으나 slow 행동은 소수의 major gene에 의하여 지배되는 것을 알았다.

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복제수 증폭시스템과 염색체 분단기술을 이용한 Simultaneous YAC Manipulation-Amplification (SYMA) 시스템의 개발 (Development of Simultaneous YAC Manipulation-Amplification (SYMA) system by Chromosome Splitting Technique Harboring Copy Number Amplification System)

  • 김연희;남수완
    • 생명과학회지
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    • 제20권5호
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    • pp.789-793
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    • 2010
  • 복잡한 진핵생물에서의 물리적 지도 작성이나 기능해석에 효모인공염색체(YAC)를 이용하기 위해서는 원하는 target region의 인공염색체화 및 single-copy인 YAC의 복제수를 늘이는 것이 요구된다. 본 연구에서는 YAC manipulation system에 복제수 증폭시스템(copy number amplification system)을 도입한 Simultaneous YAC Manipulation-Amplification (SYMA) system을 구축하였다. 식물염색체를 가진 YAC clone의 splitting과 증폭을 위해 conditional centromere와 thymidine kinase (TK) 유전자를 가진 pBGTK plasmid를 구축하였고, splitting fragment의 PCR을 위한 주형으로 사용하였다. 590 kb의 YAC clone은 splitting과 동시에 copy number amplification element를 가진 100 kb YAC와 490 kb YAC로 분리되었고, 100 kb YAC는 유도기질로 3 mg/ml sulfanilamide와 $50\;{\mu}g/ml$ methotrexate (S3/M50)의 첨가에 의해 14.4배로 그 복제수가 증가하였음을 확인할 수 있었다.

Multi-Bioindicators to Assess Soil Microbial Activity in the Context of an Artificial Groundwater Recharge with Treated Wastewater: A Large-Scale Pilot Experiment

  • Michel, Caroline;Joulian, Catherine;Ollivier, Patrick;Nyteij, Audrey;Cote, Remi;Surdyk, Nicolas;Hellal, Jennifer;Casanova, Joel;Besnard, Katia;Rampnoux, Nicolas;Garrido, Francis
    • Journal of Microbiology and Biotechnology
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    • 제24권6호
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    • pp.843-853
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    • 2014
  • In the context of artificial groundwater recharge, a reactive soil column at pilot-scale (4.5 m depth and 3 m in diameter) fed by treated wastewater was designed to evaluate soil filtration ability. Here, as a part of this project, the impact of treated wastewater filtration on soil bacterial communities and the soil's biological ability for wastewater treatment as well as the relevance of the use of multi-bioindicators were studied as a function of depth and time. Biomass; bacterial 16S rRNA gene diversity fingerprints; potential nitrifying, denitrifying, and sulfate-reducing activities; and functional gene (amo, nir, nar, and dsr) detection were analyzed to highlight the real and potential microbial activity and diversity within the soil column. These bioindicators show that topsoil (0 to 20 cm depth) was the more active and the more impacted by treated wastewater filtration. Nitrification was the main activity in the pilot. No sulfate-reducing activity or dsr genes were detected during the first 6 months of wastewater application. Denitrification was also absent, but genes of denitrifying bacteria were detected, suggesting that the denitrifying process may occur rapidly if adequate chemical conditions are favored within the soil column. Results also underline that a dry period (20 days without any wastewater supply) significantly impacted soil bacterial diversity, leading to a decrease of enzyme activities and biomass. Finally, our work shows that treated wastewater filtration leads to a modification of the bacterial genetic and functional structures in topsoil.

Disentangling Evolutionary Pattern and Haplotype Distribution of Starch Synthase III-1 (SSIIIb) in Korean Rice Collection

  • Bhagwat Nawade ;Yong-Jin Park
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.214-214
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    • 2022
  • Soluble starch synthases (SSs) elongate α-glucans from ADP-Glc to the glucan nonreducing ends and play a critical role in synthesizing resistant starch in the rice. A total of 10 SSs isoforms were reported in rice, including granules-bound starch synthase I (GBSSI), GBSSII, starch synthase I (SSI), SSIIa (SSII-3), SSIIb (SSII-2), SSIIc (SSII-1), SSIIIa (SSIII-2), SSIIIb (SSIII-1), SSIVa (SSIV-1), and SSIVb (SSIV-2). SSIII proteins are involved in forming the B chain and elongating cluster filling chains in amylopectin metabolism. The functions of SSIIIb (SSIII-1) are less clear as compared to SSs. Here, we sought to shed light on the genetic diversity profiling of the SSIII-1 gene in 374 rice accessions composed of 54 wild-type accessions and 320 bred cultivars (temperate japonica, indica, tropical japonica, aus, aromatic, and admixture). In total, 17 haplotypes were identified in the SSIII-1 coding region of 320 bred cultivars, while 44 haplotypes were detected from 54 wild-type accessions. The genetic diversity indices revealed the most negative Tajima's D value in the temperate-japonica, followed by the wild type, while Tajima's D values in other ecotypes were positive, indicating balancing selection. Nucleotide diversity in the SSIII-1 region was highest in the wild group (0.0047) while lowest in temperate-japonica. Lower nucleotide diversity in the temperate-japonica is evidenced by the negative Tajima's D and suggested purifying selection. The fixation index (FST) revealed a very high level of gene flow (low FST) between the tropical-japonica and admixture groups (FST=-0.21) followed by admixture and wild groups (-0.04), indica and admixture groups (0.02), while low gene flow with higher FST estimates between the temperate-japonica and aus groups (0.72), tropical-japonica and aromatic groups (0.71), and temperate-japonica and admixture groups (0.52). Taken together, our study offers insights into haplotype diversity and evolutionary fingerprints of SSIII-1. It provides genomic information to increase the resistant starch content of cooked rice.

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