• Title/Summary/Keyword: Gene-gun

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Food Fraud Monitoring of Commercial Sciaenidae Seafood Product Using DNA Barcode Information (DNA barcode를 이용한 민어과 수산가공품 진위판별 모니터링)

  • Park, Eun-Ji;Jo, Ah-Hyeon;Kang, Ju-Yeong;Lee, Han-Cheol;Park, Min-Ji;Yang, Ji-Young;Shin, Ji-Young;Kim, Gun-Do;Kim, Jong-Oh;Seo, Yong-Bae;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.574-580
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    • 2020
  • In this study we sought to determine the food fraud by discriminating species of commercial seafood product such as Larimichthys polyactis, Larimichthys crocea, Pennahia argentatus, and Miichthys miiuy, which are difficult to morphologically discriminate. After amplifying the mitochondrial cytochrome c oxidase subunit I gene of the reference fish, the DNA sequences of the amplified PCR products were analyzed. As a result, a 655 bp sequence for species identification was selected for use as DNA barcodes. To confirm the DNA data and primer set, the DNA barcode sequence of each fish was compared to that in that in the NCBI. All of the DNA barcode data were matched with the gene sequence of each fish in the NCBI. A total of 32 processed seafood products (8 L. polyactis, 12 L. crocea, 3 Pennahia argentatus, and 9 Miichthys miiuy) were investigated. Homology of 97% or more in DNA sequences was judged as the same species. As a result of the monitoring, there were no discovered cases of forgery or alteration. However, the use of a raw material name having no matching standard name in the Korea Food Code may cause consumer confusion. Therefore, it is suggested that the standard name or scientific name be co-labeled with the raw material name on seafood products to prevent consumer confusion.

Phylogenetic Analysis on Wild Cordyceps Collected from Miryang Region of South Korea (밀양근교에서 채집한 야생 동충하초 계통의 PCR 산물에 근거한 계통 유전학적 연구)

  • Park, Hyeancheal;Lee, Sangmong;Park, Namsook
    • Korean Journal of Plant Resources
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    • v.34 no.1
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    • pp.1-16
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    • 2021
  • The phylogenetic relationships among thirty-two strains (P1~P32; including Cordyceps sp., Paecilomyces sp., Beauveria sp., Aranthomyces sp., Isaria sp. and Himenostilbe sp.) in Miryang region located in the southern part of Korea, were investigated based on internal transcribed spacer (ITS) sequences of ribosomal DNA. A fragment of ITS region was amplified by polymerase chain reaction (PCR) using the specific primer pairs ITS1 and ITS4. After obtained same size of PCR products from various strains, we cloned them into a pGEM-T easy vector to determine their sequences. BLAST analyses of the nucleotide sequence ITS1, 5.8S and ITS2 gene fragments revealed the identity and their phylogenetic relationship. Among 32 strains isolated from Miryang region, Cordyceps militaris was shared 100% sequences with Genbank (AY49191, EU825999, AY491992), while some species are not shared perfectly with reported sequences. For example, strain P17 (P. tenuipes in Ulju-gun Gaji Mountain) has some differences among the other strains of P. tenuipes (Miryang-si Jocheon-eup, Miryang-si Gaji Mountain) and those of gene bank. We conclude that ITS analyses with strains in the suburbs of Miryang in this study can be effectively used as a tool for classification, evaluation and collection of the natural eco-type genetic resources.

Development and Validation of Multiplex Polymerase Chain Reaction to Determine Squid Species Based on 16s rRNA Gene (오징어류 종 판별을 위한 다중 유전자 검사법 개발 및 검증)

  • Kim, Hyunsu;Seo, Yong Bae;Choi, Seong-Seok;Kim, Jin-Hee;Shin, Jiyoung;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Food Hygiene and Safety
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    • v.30 no.1
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    • pp.43-50
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    • 2015
  • In this study, single PCR and multiplex PCR tests were examined for identification of four types of squid species (giant squid, cuttlefish, octopus, beka squid) purchased from fish market as well as aquatic processed products in Busan. To design the specific primers against each species, the nucleotide sequences of the mitochondrial 16s rRNA gene of Architeuthis dux, Todarodes pacificus, Enteroctopus dofleini, Enteroctopus megalocyathus, Uroteuthis chinensis, Uroteuthis duvauceli, Uroteuthis edulis groups were analyzed for the identification of each species registered in the GeneBank (www.ncbi.nlm.nih.gov) and have been used for comparative analysis. In order to obtain the size variation of amplified fragments on multiplex PCR, we designed KOJ-F, OJ-F, OCT-F, HAN-F, ALLR primers for each species. The optimal PCR conditions and primers were selected for four types of squid species to determine target base sequences in its PCR products. In the case of single PCR, giant squid was only amplified by KOJ-F/ALLR primer; cuttlefish was only amplified by OJ-F/ALLR primer; octopus was only amplified by OCT-F/ALLR primer; and beka squid was only amplified by HAN-F/ALLR primer. For multiplex PCR, the mixture of four kinds of genomic DNA (giant squid, cuttlefish, octopus, beka squid) been prepared as a template and used together with the mixture of KOJ-F/OJ-F/OCT-F/HAN-F/ALLR primers in the reaction. By the multiplex PCR, it is confirmed that four samples are correspond to multiple simultaneous amplicon. Finally, we validated the established methods of multiplex PCR in the aquatic processed products. Although the mitochondrial 16s rRNA primers used in this study was useful as a marker for detection of each species among them, the study indicated that the established multiplex PCR method can be more useful tool for monitoring the processed products.

Optimization of particle gun-mediated transformation system in Cymbidium (유전자총을 이용한 형질전환 심비디움 식물체 생산체계 최적화)

  • Noh, Hee-Sun;Kim, Mi-Seon;Lee, Yu-Mi;Lee, Yi-Rae;Lee, Sang-Il;Kim, Jong-Bo
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.293-300
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    • 2011
  • This study is conducted to develop an efficient transformation system via particle bombardment with PLBs (Protocorm-like bodies) in Cymbidium. For this, pCAMBIA3301 vector which carries a herbicide-resistant bar gene and gus gene as a reporter gene was used for transformation with Cymbidium cultivars 'Youngflower ${\times}$ masako' line. To select transformants, proper concentration of herbicide, PPT (phosphinotricin), should be determined. As a result, 5 mg/l of PPT was selected as a proper concentration. Further, proper conditions for particle bombardment were determined to obtain a high frequency of transformation. Results showed that 1.0 ${\mu}g$ of DNA concentration, 1,100 and 1,350 psi for helium gas pressure, 1.0 ${\mu}m$ of gold particle and 6 cm of target distance showed the best result for the particle bombardment experiment. Also, pre-treatment with combination 0.2 M sorbitol and 0.2 M mannitol for 4 hrs prior to genetic transformation increased the transformation efficiency up to 2.5 times. Using transformation system developed in this study, 3.2 ~ 4.0 transgenic cymbidium plants can be produced from 100 bombarded PLBs on average. Putative transgenic plants produced in this system confirmed the presence of the bar gene by PCR analysis. Also, leaves from randomely selected five transgenic lines were applied for Basta solution (0.5% v/v) to check the resistance to the PPT herbicide. As a result, three of them showed resistance and one of them showed the strongest resistance with the maintenance of green color as non-transformed plants showed. Using this established transformation system, more genes of interests can be introduced into Cymbidium plants by genetic transformation in the future.

Analysis of Major Foodborne Pathogens in Various Foods in Korea

  • Kim, Mi-Gyeong;Oh, Mi-Hwa;Lee, Gun-Young;Hwang, In-Gyun;Kwak, Hyo-Sun;Kang, Yun-Sook;Koh, Young-Ho;Jun, Hong-Ki;Kwon, Ki-Sung
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.483-488
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    • 2008
  • Foodborne pathogenic bacteria in various food samples in Korea were monitored and the obtained data was statistically analyzed. A total of 1,240 food samples including 280 sashimi, 244 processed frozen products, 258 kimbab (cooked rice wrapped with seaweed), 337 soybean pastes were obtained from 7 cities including Seoul in Korea. Microorganisms tested were Bacillus cereus, Salmonella spp., Staphylococcus aureus, Escherichia coli, E. coli O157:H7, Vibrio parahaemolyticus, Yersinia enterocolitica, Listeria monocytogenes, Campylobacter jejuni, and Clostridium perfringens. The contaminated microorganisms in food samples were comprised of 10.55% B. cereus, 2.7% S. aureus, 2.0% V. parahaemolyticus, 0.8% C. perfringens, 0.2% Y. enterocolitica, and 0.1% of L. monocytogenes, respectively. Salmonella spp., C. jejuni, and E. coli O157:H7 were not detected in any of the food samples. Particularly, B. cereus that harbors the enterotoxin gene was detected in various foods and regions in Korea, therefore it should be a given special consideration not to allow the hazardous level of contamination.

Single-dose oral toxicity study of genetically modified silkworm expressing EGFP protein in ICR mouse

  • Jang, Kyung-Min;Kim, Sung-Gun;Park, Ji-Young;Choi, Won-Ho;Lee, Jae-Woo;Jegal, Hyeon-Young;Kweon, Soon-Jong;Choi, Kwang-Ho;Park, Jung-Ho
    • Korean Journal of Agricultural Science
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    • v.43 no.1
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    • pp.109-115
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    • 2016
  • Silk has had a reputation as a luxurious and sensuous fabric but it is not popular due to the expensive price and poor durability. To develop the silk materials that apply the various industries, the artificially synthesized gene can be introduced into the silkworm and expressed in the silk gland. Transgenic silkworms for the mass production of green fluorescent silks are generated using a fibroin H-chain expression system. For commercial use, safety assessment of the transgenic silkworms is essential. The purpose of this study was to examine the potential acute oral toxicity of EGFP protein expressed in genetically modified (GM) fluorescence silkworm and to obtain the approximative lethal dose in the male and female at 6-weeks ICR mice. EGFP protein was fed at a dose of 2,000 mg/kg body weight in five male or five female mice. Mortalities, clinical findings and body weight changes were monitored for 1, 3, 7, 14 days after dosing. At the end of 14 day observation period, all mice were sacrificed, and the postmortem necropsy were performed. The test group was not observed death case. Also the effect was not admitted by test substance administration in common symptoms, the body weight and postmortem. The results of single-dose oral toxicity test showed that approximative lethal dose of EGFP protein expressed in fluorescence silkworm was considered to exceed the 2,000 mg/kg body weight in both sexes.

Stimulation of the milk protein production in MAC-T cells by purified bee venom (정제봉독의 MAC-T 세포에서 유단백 합성 촉진효과)

  • Han, Sang-Mi;Woo, Soon-Ok;Kim, Se-Gun;Jang, Hye-Ri
    • Korean Journal of Veterinary Service
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    • v.41 no.3
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    • pp.171-177
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    • 2018
  • Purified bee venom was collected from colonies of honeybees (Apis mellifera L.) using a bee venom collector under sterile conditions and then purified under strict laboratory conditions. Purified bee venom contained $63.9{\pm}5.4%$ melittin, $10.9{\pm}1.6%$ phospholipase A2, and $2.3{\pm}0.3%$ apamin. Purified bee venom has various anti-bacterial, anti-inflammatory and immunostimulating effects. In this study, we evaluated purified bee venom which are mammary gland cells, MAC-T cells are used to increase the synthesis of milk protein. Purified bee venom promoted the proliferation of MAC-T cells at concentrations below $1{\mu}g/mL$, but cytotoxicity at $10{\mu}g/mL$ and above. As a result of the increase in the synthesis of ${\beta}-casein$, a milk protein after treatment with MAC-T cells at a concentration of the bee venom without cytotoxicity, the ${\beta}-casein$ content in the cell culture was increased when treated at a concentration of 1 ng/mL or more. In addition, it was confirmed that purified bee venom significantly increased the expression of bovine ${\beta}-casein$ (bCSNB) mRNA, a ${\beta}-casein$ synthesis gene, at a concentration of 1 ng/mL or more. These results suggest that purified bee venom can be used to increase the production of livestock by ultimately increasing the expression of milk protein.

Pharmacological Systemic Analysis of Curcumae Radix in Lipid Metabolism (시스템 분석을 통한 지질대사에서 울금의 약리작용)

  • Jo, Han Byeol;Kim, Ji Young;Kim, Min Sung;An, Won Gun;Lee, Jang-Cheon
    • Herbal Formula Science
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    • v.26 no.3
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    • pp.237-250
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    • 2018
  • Objectives : This study is a pharmacological network approach, aimed to identify the potential active compounds contained in Curcumae Radix, and their associated targets, to predict the various bio-reactions involved, and finally to establish the cornerstone for the deep-depth study of the representative mechanisms. Methods : The active compounds of Curcumae Radix have been identified using Traditional Chinese Medicine System Pharmacology Database and Analysis Platform. The UniProt database was used to collect each of information of all target proteins associated with the active compounds. To find the bio-metabolic processes associated with each target, the DAVID6.8 Gene Functional classifier tool was used. Compound-Target and Target-Pathway networks were analyzed via Cytoscape 3.40. Results : The target information from 32 potential active compounds of Curcumae Radix was collected through TCMSP analysis. The active compounds interact with 133 target genes engaging in total of 885 biological pathways. The most relevant pathway was the lipid-related metabolism, in which 3 representative active compounds were naringenin, oleic acid, and ${\beta}-sitosterol$. The mostly targeted proteins in the lipid pathway were ApoB, AKT1 and PPAR. Conclusions : The pharmacological network analysis is convenient approach to predict the overall metabolic mechanisms in medicinal herb research, which can reduce the processes of various experimental trial and error and provide key clues that can be used to validate and experimentally verify the core compounds.

Human Embryonic Stem Cells Experience a Typical Apoptotic Process upon Oxidative Stress

  • Lee, Gun-Soup;Lee, Young-Jae;Kim, Eun-Young;Park, Se-Pill;Lim, Jin-Ho
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.97-97
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    • 2003
  • Embryonic stem (ES) cells, derived from preimplantation embryos, are able to differentiate into various types of cells consisting the whole body, or pluripotency. In addition to the plasticity, ES cells are expected to be different from terminally differentiated cells in very many ways, such as patterns of gene expressions, ability and response of the cells in confronting environmental stimulations, metabolism, and growth rate. As a model system to differentiate these two types of cells, human ES (hES, MB03) cells and terminally differentiated cells (HeLa), we examined the ability of these two types of cells in confronting a severe oxidative insult, that is $H_2 O_2$. Ratio of dying cells as determined by the relative amount of dye neutral red entrapped within the cells after the exposures. Cell death rates were not significantly different when either MB03 or HeLa were exposed up to 0.4 mM $H_2 O_2$. However, relative amount of dye entrapped within the cells sharply decreased down to 0.12% in HeLa cells when the cells were exposed to 0.8 mM $H_2 O_2$, while it was approximately 54% in MB03. Pretreatment of cells with BSO (GSH chelator) and measurement of GSH content results suggest that cellular GSH is the major defensive mechanism of hES cells. Induction of apoptosis in hES cell was confirmed by DNA laddering, induction of Bax, and chromatin condensation. In summary, hES cells 1) are extremely resistant to oxidative stress, 2) utilize GSH as a major defensive mechanism. and 3) experience apoptosis upon exposure to oxidative stress.

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Control Effect of Stenotrophomonas maltophilia BW-13 strain to the lettuce Bottom rot

  • Park, Jong-Young;Kim, Hyun-Ju;Bak, Joung-Woo;Lee, Kwang-Youll;Jun, Ok-Ju;Lee, Jin-Woo;Jung, Soon-Je;Moon, Byung-Ju
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.103.1-103
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    • 2003
  • An antagonistic bacteria, Stenotrophomonas maitophilia BW-13 strain which was effectively inhibited mycerial growth of Bottom rot pathogen, Rhizoctonia solani PY-1 strain was isolated from the rhizosphere of the lettuce in Uiryeong-Gun, Gyeongsangnam-Do from 2002 to 2003. For the biological control, the most suitable inoculum and its density of pathogen, PY-1 strain ware tested prior biological control test, For the pathogenicity test, A inoculum (wheat bran)sawdust+rice bran+PDB) showing disease incidence of 100% was selected as the most suitable inoculum, which showed more effective than B inoculum (sawdust+rice bran+DW) and mycelial disc. also, In selection of the amount of inoculum (40g, 50g, 60g, 70g, 80g), most suitable amount of inoculum of pathogen determined as 40g showing disease incidence of 80%. For the selection of effective microorganism to control bottom rot on lettuce, about 200 isolates were isolated from the diseased soil and lettuce leaves, and examined their antifungal activity to the pathogen on PDA. As the pots assay, BW-13 strain showed the highest control value as 90%, and followed by R-13 and R-26 strain as 80% and 60%, respectively. Selected BW-13 isolates identified as 5. maltophilia (GeneBank accession no. AJ293473.1, 99%) by 16S rRNA sequencing. This is the first report on the biological control using by S. maltophilia to the bottom rot pathogen, Rhizoctonia solani PY-1 strain.

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