• Title/Summary/Keyword: Synthetic biology

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Current Status and Future Perspectives of Natural Enemy Research in Korea: Analysis of Research Papers Published in the Korean Journal of Applied Entomology from 1990 to 2020 (우리나라 천적 연구의 현황 및 미래 전략: 1990년부터 2020년까지 한국응용곤충학회지 게재 논문 분석)

  • Cho, Jum Rae;Kim, Jeong Hwan;Seo, Meeja;Choi, Seon U;Lim, Un Taek;Lee, Kyeongyeoll
    • Korean journal of applied entomology
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    • v.60 no.3
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    • pp.287-304
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    • 2021
  • Since 1995, researches on natural enemies have been conducted extensively in Korea. Research papers on natural enemies published in the Korean Journal of Applied Entomology from 1990 to 2020 were 130, which is 8.4% of all published papers during the same period. In 1990s, most research papers study the searching and biological characteristics of natural enemies, whereas the proportion of research papers related to the field application using the developed natural enemies has been increasing since 2010s. A total of 37 excellent natural enemies have been developed including 24 indigenous and 13 introduced natural enemies. In addition, 28 kinds of booklets and/or manuals were developed for field application of natural enemies. Although successes in research and development have been achieved since that period, more researches on search for and/or introduction of excellent natural enemy suitable for the Korean cultivation environment, mass production technology that can reduce cost, and quality control program in producing and distributing natural enemy remain to be pursued in the future. Furthermore, there is a need to develop the technology that can be used in compatible way with natural enemies and other crop protection agents including synthetic insecticides.

Determination of the Length of Target Recognition Sequence in sgRNA Required for CRISPR Interference (CRISPR 간섭에 필요한 sgRNA 표적 인식 서열 길이의 결정)

  • Kim, Bumjoon;Kim, Byeong Chan;Lee, Ho Joung;Lee, Sang Jun
    • Microbiology and Biotechnology Letters
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    • v.49 no.4
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    • pp.534-542
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    • 2021
  • Single-molecular guide RNA (sgRNA) plays a role in recognizing the DNA target sequence in CRISPR technology for genome editing and gene expression control. In this study, we systematically compared the length of the target recognition sequence in sgRNAs required for genome editing using Cas9-NG (an engineered Cas9 recognizing 5'-NG as PAM sequence) and gene expression control using deactivated Cas9-NG (dCas9-NG) by targeting the gal promoter in E. coli. In the case of genome editing, the truncation of three nucleotides in the target recognition sequence (TRS) of sgRNA was allowed. In gene expression regulation, we observed that target recognition and binding were possible even if eleven nucleotides were deleted from twenty nucleotides of the TRS. When 4 or more nucleotides are truncated in the TRS of the sgRNA, it is thought that the sgRNA/Cas9-NG complex can specifically bind to the target DNA sequence, but lacks endonuclease activity to perform genome editing. Our study will be helpful in the development of artificial transcription factors and various CRISPR technologies in the field of synthetic biology.

Detection of Microcystin Synthetic Cyanobacteria and Variation of Intracellular Microcystin Synthesis Using by eDNA and eRNA in Freshwater Ecocystem (담수환경에서 eDNA와 eRNA를 이용한 Microcystin 합성 남조류 탐색 및 세포 내 Microcystin 생합성 활성 변화)

  • Keonhee Kim;Chaehong Park;Hyeonjin Cho;Daeryul Kwon;Soon-Jin Hwang
    • Korean Journal of Ecology and Environment
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    • v.56 no.1
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    • pp.1-13
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    • 2023
  • Targeting Microcystin (MC), which is most abundantly detected in the North-Han River water area, we analyzed the relationship between the MC biosynthesis gene (mcyA gene), cyanobacteria cell density, and MC concentration, derived an RNA-MC conversion formula, and derived the cyanobacteria. The concentration of MC present in cells was predicted. In the North-Han River waters, the mcyA gene was found mainly at downstream sites of the North-Han River after Muk-Hyeon Stream junction, and higher copy numbers were found on average than other sites. In the Uiam Lake waters upstream of the North-Han River, the mcyA gene copy number increased at the Kong-Ji Stream point, and after September, the mcyA gene copy number decreased throughout the North-Han River waters. The expression of the mcyA gene was concentrated in the short period of summer due to the spatio-temporal difference between upstream and downstream water bodies. The mcyA gene expression level was not only highly correlated with MC concentration, but also correlated with the cell density of Microcystis aeruginosa and Dolichospermum circinale, which are known to biosynthesize MC. Six conversion formulas derived based on the RNA-MC relationship showed statistical significance (p<0.05) and exhibited high correlation coefficients (r) of 0.9 or higher. The expression level of MC biosynthesis gene present in eRNA determines the synthesis of cyanotoxin substances in water, quickly quantifies gene activity, and can be fully utilized for early warning of MC development.

The effects of microplastics on marine ecosystem and future research directions (미세플라스틱의 해양 생태계에 대한 영향과 향후 연구 방향)

  • Kim, Kanghee;Hwang, Junghye;Choi, Jin Soo;Heo, Yunwi;Park, June-Woo
    • Korean Journal of Environmental Biology
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    • v.37 no.4
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    • pp.625-639
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    • 2019
  • Microplastics are one of the substances threatening the marine ecosystem. Here, we summarize the status of research on the effect of microplastics on marine life and suggest future research directions. Microplastics are synthetic polymeric compounds smaller than 5 mm and these materials released into the environment are not only physically small but do not decompose over time. Thus, they accumulate extensively on land, from the coast to the sea, and from the surface to the deep sea. Microplastic can be ingested and accumulated in marine life. Furthermore, the elution of chemicals added to plastic represents another risk. Microplastics accumulated in the ocean affect the growth, development, behavior, reproduction, and death of marine life. However, the properties of microplastics vary widely in size, material, shape, and other aspects and toxicity tests conducted on several properties of microplastics cannot represent the hazards of all other microplastics. It is necessary to evaluate the risks according to the types of microplastic, but due to their variety and the lack of uniformity in research results, it is difficult to compare and analyze the results of previous studies. Therefore, it is necessary to derive a standard test method to estimate the biological risk from different types of microplastics. In addition, while most of the previous studies were conducted mostly on spheres for the convenience of the experiments, they do not properly reflect the reality that fibers and fragments are the main forms of microplastics in the marine environment and in fish and shellfish. Furthermore, studies have been conducted on additives and POPs (persistent organic pollutants) in plastics, but little is known about their toxic effects on the body. The effects of microplastics on the marine ecosystems and humans could be identified in more detail if standard testing methods are developed, microplastics in the form of fibers and fragments rather than spheres are tested, and additives and POPs are analyzed. These investigations will allow us to identify the impact of microplastics on marine ecosystems and humans in more detail.

Detection of Auxotrophic Mutants form Valsa ceratosperma, the Causal Fungus of Apple Canker (사과나무 부란병균(腐爛病菌) Valsa ceratosperma에서의 Auxotrophic Mutants의 검출(檢出))

  • Hong, Yeon Gyu;Uhm, Jae Youl
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.119-126
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    • 1987
  • This study was conducted to elucidate the most appropriate method to obtain auxotrophic mutants from Valsa ceratosperma, the causal fungus of apple canker, which may be used as a gene marker in detecting the transfer of the factors of avirulent strains to virulent strains. Among the 3 kinds of synthetic media tested, each have two formula for minimal and complete, the medium which has been used in study of Endothia parasitica (E. P medium) was turned out to be most appropriate for the growth of V. ceratosperma. A medium for single colony formation from pycnidiospore of this fungus was developed by adding 0.5% L - sorbose to the E. P minimal medium. The period of incubation in dark for preventing the photoreactivation after U. V irradiation was estimated as about 60hrs at which most of the spores become binucleate. Largest number of putative auxotrophs were obtained at about 50second of irradiation to the spores smeared on the medium for single colony formation, at which the survival rate of spores was 5 to 6 percent. With these method developed in this experiment, 161 isolates of putative auxotrophs were detected among which the nutrient requirement for 10 isolates were determined. Five out of 10 mutants were still virulent to apple tree and all but one could not sporulate.

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Comparison of AndroMed and Triladyl Extender for Freezing of Jeju Native Black Bull Semen (제주흑우 정액 동결을 위한 AndreoMed와 Triladyl 희석제의 동결성 비교)

  • Cho, Sang-Rae;Cho, In-Chel;Han, Sang-Hyun;Kang, Tae-Young;Cho, Won-Mo;Chae, Hyun-Seok;Kim, Nam-Young;Park, Yong-Sang;Kang, Yong-Jun;Kim, Young-Hoon;Ko, Yeoung-Gyu;Kim, Hyun-Jong;Ko, Moon-Suck
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.155-159
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    • 2013
  • This study was carried out to investigate synthetic extender for semen cryopreservation of Jeju Native Black Bull. The semen was collected using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by Tris-Egg yolk extender and contrifuged in 1,500 rpm for 15 minutes. The supernatant was removed. The pellect was diluted to final sperm concentration of $2{\times}10^8/ml$ by doubling in every 30 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 4 hours at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm for 10 minutes. The height and duration affect the freezing speed by temperature. The frozen straw was plunged to $LN_2$. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. Frozen-thawed sperm were evaluated sperm viability, membrane integrity and acrosome integrity. Post-thawed sperm viability has been significantly higher (p<0.05) in fresh sperm ($93.27{\pm}1.62%$) than frozen-thawed sperm ($73.34{\pm}3.27%$). However, there were no significant differences between fresh and frozen-thawed dead cell rate ($7.35{\pm}2.63$ vs, $13.71{\pm}2.85$). In sperm motility, between Triladyl and AndroMed Extender, there was no significant different ($72.86{\pm}2.83$ vs, $81.47{\pm}2.48$), similarly, the dead cell rates was similar ($18.41{\pm}3.42%$ and $17.26{\pm}4.25$). The results of our study suggest that AndroMed to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Jeju Native Black bull semen.

cDNA Cloning and Stage-Dependant Expression of Arylphorin Gene from Chinese Oak Silkworm, Antheraea pernyi (작잠(Antheraea pernyi) 아릴포린(Arylphorin) 유전자의 cDNA 클로닝 및 아릴포린 유전자의 발육시기 의존성 발현양상)

  • Lee, Sang-Mong;Hwang, Jae-Sam;Park, Nam-Sook;Kim, Yong-Gyun;Kim, Keun-Ki;Son, Hong-Joo;Park, Hyun-Chul;Jin, Byung-Rae
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1193-1200
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    • 2010
  • The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

Production of Violacein by a Novel Bacterium, Massilia sp. EP15224 Strain (Violacein을 생산하는 Massilia sp. EP15224 균주)

  • Yoon, Sang-Hong;Baek, Hee-Jin;Kwon, Soon-Wu;Lee, Chang-Muk;Sim, Joon-Soo;Hahn, Bum-Soo;Koo, Bon-Sung
    • Microbiology and Biotechnology Letters
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    • v.42 no.4
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    • pp.317-323
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    • 2014
  • Violacein has received much attention due to its various important biological activities, including broad-spectrum antibacterial and antifungal activity, anti-malarial, anti-tumoral, anti-oxidant, and anti-diarrheal activities. EP15224 strain isolated from forest soils in Korea was found to be a new species belonged to the genus Massilia based on its 16S ribosomal DNA sequences. The 16S ribosomal DNA of strain EP15224 displayed 97% homology with Massilia sp. BS-1, the nearest violacein-producing bacterium. Strain EP15224 produced bluish-purple pigment well in a synthetic MM2 medium containing glucose, $(NH_4)_2SO_4$, $Na_2HPO_4{\cdot}7H_2O$, $KH_2PO_4$, $MgSO_4{\cdot}7H_2O$, and 1 mM $\small{L}$-tryptophan. The chemical analysis of the pigment by LC/MS/MS showed that it is violacein with molecular weight of 343.34. This is the second report on the production of violacein by a Massilia species. In this study, the optimal culture conditions for violacein production were established under which 280 mg/l crude violacein was produced : glucose 2 g/l, $(NH_4)_2SO_4$ 1 g/l, $Na_2HPO_4{\cdot}7H_2O$ 2 g/l, $KH_2PO_4$ 1 g/l, $MgSO_4{\cdot}7H_2O$ 0.1 g/l, L-tryptophan 0.24 g/l, 25 ml medium in a 250 ml flask, with an inoculumn size of 10% (v/v), 72 h of cultivation with 250 rpm at $25^{\circ}C$.