• Title/Summary/Keyword: gel electrophoresis

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Powdery Mildew Resistance Phenotype Test & Genotype Test in C. moschata

  • Jong-Gyu Park
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.290-290
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    • 2022
  • Powdery mildew is known to be one of the serious diseases in C. moschata cultivation. Plants infected with powdery mildew cause damage to cultivation areas such as occurrence of deformity fruit and decrease in quantity. also, it has been reported that many farms have difficulties in controlling powdery mildew due to the outbreak under various conditions throughout the year. Therefore, this study intends to perform a phenotype test and a genotype test for C. moschata 60 lines grown in Jenong S&T. Podospareaxanthii, known as a pathogen that causes powder mildew disease in pumpkins in Korea, was collected and used as an inoculation source, phenotype test was performed by examining the infection area rate(%) of powdery mildew disease that occurred in leaves 25 days after inoculation. It was determined that 0% of the infection area rate was in the first stage, 1 to 5% in the second stage, 6 to 15% in the third stage, 16 to 30% in the fourth stage, and 31% or more in the fifth stage, The first and second stages were judged as resistance, the third as moderate resistance, and the fourth and fifth stages as sensitivity. As a result of the phenotype test, it was confirmed that the resistance was 21 points, moderate resistance was 14 points, and sensitivity was 25 points. After searching for the genes related to powdery mildew resistance resistance, pm-0, CmbHLH87, and LOC111453072, 21 points of resistance and 9 points of moderate resistance identified through phenotype tests were identified through gel electrophoresis after polymerase chain reaction(PCR) using 5 primers related to 3 genes. As a result of genotype testing of a total 30 points, the CmbHLH87 and LOC111453072 gene were found to be resistant bands in all points, PMR1 was identified as 20 points for resistance, 4 points for moderate resistance, and 6 points for sensitivity, PMR2 was not identified in the entire band, and PMR5 was identified as 18 point for resistance, 3 points for moderate resistance, and 9 points for sensitivity. As a result, when comparing the phenotype test results and genotype test results, CmbHLH87 and LOC111453072 genes was 100% consistent in resistance and moderate resistance, PMR1 was 95.2% in resistance, 44.4% in moderate resistance, and PMR5 was 90% in resistance and 33.3% in moderate resistance, PMR2 was not consistent in resistance and moderate resistance. Therefore, it is expected that more accurate PMR test will be possible by using molecular markers(PMR1, PMR5) and by developing CmbHLH87 and LOC111453072 gene-related molecular markers.

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Prevalence of Toxin Genes and Antibiotic Resistance Profiles of Vibrio vulnificus strains isolated from Jeju Island (제주도에서 분리된 비브리오패혈증균의 독소 유전자 분포 및 항생제 내성)

  • Eunok Kang;Man Jae Cho;Ye-Seul Heo;Eun A Koh
    • Journal of Food Hygiene and Safety
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    • v.38 no.5
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    • pp.381-389
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    • 2023
  • Vibrio vulnificus, the most fatal waterborne and foodborne pathogens of 50% fatality rate in the world, is common in seawater and occurs particularly in warmer months. In this study, we investigated the toxin genes using reverse transcription-polymerase chain reaction (RT-PCR), antibiotic resistance status using Vitek, and genetic characteristics using pulsed-field gel electrophoresis (PFGE) of different V. vulnificus strains isolated from the Jeju Island seawater, distribution fishery products, and fish tanks. We examined a total of 487 samples and isolated a total of 46 strains (including overlapping strains) of V. vulnificus, 44 strains from seawater and 1 strain each from fishery products and fish tank. We detected toxin gene vvhA in all 46 strains and rtxA, viu in 8 strains (17.4%) and 9 strains (19.6%) strains, respectively. Antibiotic resistance tests indicated 100% resistance to cefoxitin antibiotics. The PFGE analysis of the 46 strains identified a total of 6 types showed 100% homology and the degree of similarity was 81.3-98.0%; however, there were no similarity between the regions and samples. These results indicate that V. vulnificus isolated from the seawater, fishery products, and fish tanks should be continuously monitored as cases of food poisoning caused by V. vulnificus with toxin genes have been reported in Jeju Island.

Exploration of nutritional and bioactive peptide properties in goat meat from various primal cuts during in vitro gastrointestinal digestion and absorption

  • Pichitpon Luasiri;Papungkorn Sangsawad;Jaksuma Pongsetkul;Pramote Paengkoum;Chatsirin Nakharuthai;Saranya Suwanangul;Sasikan Katemala;Narathip Sujinda;Jukkrapong Pinyo;Jarunan Chainam;Chompoonuch Khongla;Supaluk Sorapukdee
    • Animal Bioscience
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    • v.37 no.6
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    • pp.1096-1109
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    • 2024
  • Objective: This research aims to explore the nutritional and bioactive peptide properties of goat meat taken from various primal cuts, including the breast, shoulder, rib, loin, and leg, to produce these bioactive peptides during in vitro gastrointestinal (GI) digestion and absorption. Methods: The goat meat from various primal cuts was obtained from Boer goats with an average carcass weight of 30±2 kg. The meat was collected within 3 h after slaughter and was stored at -80℃ until analysis. A comprehensive assessment encompassed various aspects, including the chemical composition, cooking properties, in vitro GI digestion, bioactive characteristics, and the bioavailability of the resulting peptides. Results: The findings indicate that the loin muscles contain the highest protein and essential amino acid composition. When the meats were cooked at 70℃ for 30 min, they exhibited distinct protein compositions and quantities in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis profile, suggesting they served as different protein substrates during GI digestion. Subsequent in vitro simulated GI digestion revealed that the cooked shoulder and loin underwent the most significant hydrolysis during the intestinal phase, resulting in the strongest angiotensin-converting enzyme (ACE) and dipeptidyl peptidase-IV (DPP-IV) inhibition. Following in vitro GI peptide absorption using a Caco-2 cell monolayer, the GI peptide derived from the cooked loin demonstrated greater bioavailability and a higher degree of ACE and DPP-IV inhibition than the shoulder peptide. Conclusion: This study highlights the potential of goat meat, particularly cooked loin, as a functional meat source for protein, essential amino acids, and bioactive peptides during GI digestion and absorption. These peptides promise to play a role in preventing and treating metabolic diseases due to their dual inhibitory effects on ACE and DPP-IV.

Serovars and Genetic Characteristic of Salmonella spp. Isolates from Jeju Island, South Korea (제주도에서 분리된 살모넬라균의 혈청형 및 유전학적 특성)

  • Eunok Kang;Man Jae Cho;Chang Hui Yang
    • Journal of Food Hygiene and Safety
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    • v.39 no.2
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    • pp.134-151
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    • 2024
  • Salmonella spp. is among the most important water-borne and food-borne pathogens and is one of the most common causes of human gastroenteritis and diarrheal diseases globally. In this study, Salmonella spp. isolated from food, environmental samples, and patients with food poisoning or diarrhea were investigated Salmonella serovars, antibiotic resistance using Vitek2, and genetic characteristics through pulsed-field gel electrophoresis (PFGE). Salmonella spp. of 339 strains, including 26 strains from food or environmental samples and 313 strains from patients, were isolated from Jeju Island of South Korea between 2020 and 2023. The monthly number of isolated Salmonella spp. gradually increased from March, with the highest number being in August. No significant differences in Salmonella spp. isolated from patients according to gender was observed. However, Salmonella spp. was most frequently isolated from people aged 70 years or older and least frequently isolated from those between ages 10 and 19 years. Salmonella spp. isolated from food or environmental samples were distributed among eight different serovars and the main serovars were identified in the order of S. Bareilly (26.9%), S. Rissen (23.1%), and S. Thompson (19.3%). Salmonella spp. isolated from patients were distributed among 27 different serovars and the main serovars were identified in the order of S. Bareilly (31.0%), S. Typhimurium (24.6%), and S. Enteritidis (11.5%). The main cause serovars of Salmonella spp. outbreaks are S. Bareilly, S. Enteritidis, S. Thompson. Antibiotic resistance tests indicated resistance to various antibiotics and some Salmonella spp. exhibited multidrug resistance. Salmonella spp. showed various genetic correlations among the 17 serovars. These results indicate that they can be used as basic data for epidemiological investigations by predicting the appearance of Salmonella spp. and providing a scientific basis.

Analysis of the Causes of a Large Food Poisoning Outbreak Attributable to Bacillus cereus (Bacillus cereus에 의한 대규모 집단식중독 원인 분석)

  • Hyunah Lee;Youngeun Ko;Dayeon Lee;KyungA Yun;Hyeonjeung Kim;Ok Kim;Junhyuk Park
    • Journal of Food Hygiene and Safety
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    • v.39 no.2
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    • pp.102-108
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    • 2024
  • This study was performed to establish the epidemiological features of a food poisoning outbreak that occurred in the cafeteria of a company in Chungcheongnam-do Province, Korea, in October 2020, and to recommend measures to prevent similar outbreaks. Twenty-one patients with acute gastroenteritis, three food handlers, seven cooking utensils, and 12 preserved food samples were subjected to viral and bacterial analyses based on procedures described in the "Manual for Detection of Foodborne Pathogens at Outbreaks". Among 135 individuals who had been served the meals, 21 (15.6%) showed symptoms of nausea and vomiting within an hour of consuming the food. Bacillus cereus were isolated from 11 (52.4%) of the 21 patients, one food service employee, one item of cooking ware, and 12 preserved food samples. In addition, we confirmed the toxin genes CER, nheA, and entFM from the isolated B. cereus strains. Pulsed-field gel electrophoresis results indicated that all of the isolated B. cereus strains were closely related, with the exception of strains obtained from one patient and one sample of preserved food. These findings provide evidence to indicate that the isolated B. cereus originated from preserved foods and an unhygienic eating environment. This outbreak highlights that the provision of food in non-commercial food systems must be thoroughly managed. In addition, it emphasizes the necessity for the correct and timely identification of causal pathogens for tracing the cause of food poisoning outbreaks, and the need to preserve food under appropriate conditions. To prevent similar cases of food poisoning, it is necessary to investigate cases based on an epidemiological approach and share the findings.

Enzymatic preparation and antioxidant activities of protein hydrolysates derived from tuna byproducts (참치 가공부산물로부터 단백가수분해물 제조 및 항산화 활성 평가)

  • Gyu-Hyeon Park;Jeong-Min Lee;Na-Young Lim;Syng-Ook Lee
    • Food Science and Preservation
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    • v.30 no.5
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    • pp.885-895
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    • 2023
  • This study aims to investigate the production and characteristics of protein hydrolysates derived from tuna byproducts (TP) using various proteolytic enzymes and to compare the antioxidant activity of the resulting hydrolysates. The TP were subjected to enzymatic hydrolysis using five different proteases: alcalase, bromelain, flavourzyme, neutrase, and papain, and the antioxidant activities of the hydrolysates were evaluated. Subsequent analysis of the available amino group contents and sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns indicated a high degree of hydrolysis in TP after treatment with all the enzymes, except for papain. Based on the RC50 values obtained from four different antioxidant analyses, all the hydrolysates exhibited similar antioxidant activity, except for the flavourzyme hydrolysate, which showed significantly higher scavenging activity against ABTS radicals and hydrogen peroxide than the other hydrolysates. These findings suggest that protein hydrolysates derived from TP hold promise as potential sources of natural antioxidants.

Enzymatic preparation and antioxidant activities of protein hydrolysates from defatted egg yolk (탈지난황을 이용한 단백가수분해물 제조 및 항산화 활성 평가)

  • Go-Eun Ko;Na-Yeong Kwak;Ha-Eun Nam;Su-Jin Seo;Syng-Ook Lee
    • Food Science and Preservation
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    • v.31 no.3
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    • pp.444-451
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    • 2024
  • This study aimed to investigate the characteristics of protein hydrolysates derived from defatted egg yolk using various proteolytic enzymes and compare the antioxidant activity of the resulting hydrolysates. The defatted egg yolk powder was subjected to enzymatic hydrolysis using four different proteases (alcalase, bromelain, flavourzyme and neutrase), and the resulting hydrolysates were evaluated for their antioxidant properties. Through analysis of available amino group contents and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, it was observed that the defatted egg yolk powder treated with alcalase, flavourzyme, and neutrase for 12 h exhibited a high degree of hydrolysis value. Based on the RC50 values obtained from two different antioxidant analyses, all hydrolysates showed comparable antioxidant activity, except for the alcalase hydrolysate, which demonstrated notably higher scavenging activity against hydrogen peroxide than the other hydrolysates. These findings suggest the potential of protein hydrolysates from defatted egg yolk, a by-product of lecithin extraction, as natural antioxidants.

Convolution Neural Network for Prediction of DNA Length and Number of Species (DNA 길이와 혼합 종 개수 예측을 위한 합성곱 신경망)

  • Sunghee Yang;Yeone Kim;Hyomin Lee
    • Korean Chemical Engineering Research
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    • v.62 no.3
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    • pp.274-280
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    • 2024
  • Machine learning techniques utilizing neural networks have been employed in various fields such as disease gene discovery and diagnosis, drug development, and prediction of drug-induced liver injury. Disease features can be investigated by molecular information of DNA. In this study, we developed a neural network to predict the length of DNA and the number of DNA species in mixture solution which are representative molecular information of DNA. In order to address the time-consuming limitations of gel electrophoresis as conventional analysis, we analyzed the dynamic data of a microfluidic concentrating device. The dynamic data were reconstructed into a spatiotemporal map, which reduced the computational cost required for training and prediction. We employed a convolutional neural network to enhance the accuracy to analyze the spatiotemporal map. As a result, we successfully performed single DNA length prediction as single-variable regression, simultaneous prediction of multiple DNA lengths as multivariable regression, and prediction of the number of DNA species in mixture as binary classification. Additionally, based on the composition of training data, we proposed a solution to resolve the problem of prediction bias. By utilizing this study, it would be effectively performed that medical diagnosis using optical measurement such as liquid biopsy of cell-free DNA, cancer diagnosis, etc.

The relation between angiotensin converting enzyme (ACE) gene polymorphism and neonatal hyperbilirubinemia in Korea (한국인 신생아 황달과 안지오텐신 전환효소 유전자의 다형성)

  • Kim, Mi Yeoun;Lee, Jae Myoung;Kim, Ji Sook;Kim, Eun Ryoung;Lee, Hee Jae;Yoon, Seo Hyun;Chung, Joo Ho
    • Clinical and Experimental Pediatrics
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    • v.50 no.1
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    • pp.28-32
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    • 2007
  • Purpose : Human angiotensin converting enzyme (ACE) gene shows an insertion/deletion polymorphism in 16 intron, and three genotypes are determined by whether a 287 bp fragment of the DNA is present or not; II, ID and DD genotype. DD genotype has been suggested as a risk factor of chronic nephrotic disease such as IgA nephropathy and diabetic nephropathy, various cardiovascular diseases and several other diseases. ACE activity increases in acute hepatitis, chronic persistent hepatitis, chronic active hepatitis and cirrhosis. On the other hand, patients with fatty livers have normal ACE activity. This study was designed to find out the relation between polymorphsims of the ACE genes and neonatal hyperbilirubinemia in Koreans. Methods : The genomic DNA was isolated from 110 full-term Korean neonates who had hyperbilirubinemia with no obvious causes (serum bilirubin$${\geq_-}12mg/dL$$) and 164 neonates of a control population (serum bilirubin <12 mg/dL). We performed polymerase chain reaction (PCR) to see the allele of the ACE gene. Electrophoresis was done in the PCR products in 1.5 percent agarose gel, and then DNA patterns were directly visualized under ethidium bromide staining. Results : ACE genotypes in the hyperbilirubinemia group are as follows; 26.36 percent for II, 53.64 percent for ID, 20.00 percent for DD, 0.532 for I allele and 0.468 for D allele. These distributions were not significantly different from those in the control group; 24.39 percent for II, 51.83 percent for DI, 23.78 percent for DD, 0.503 for I allele and 0.497 for D allele. Conclusion : In this study, ACE gene polymorphism was detected in the neonatal hyperbilirubinemia and control group. The most frequent genotype was ID. Our results indicate that the ACE gene polymorphism is not associated with the prevalence of neonatal hyperbilirubinemia in Koreans.

Characterization of Lactate Dehydrogenase in Acanthogobius hasta (풀망둑(Acanthogobius hasta) 젖산탈수소효소의 특성)

  • Yum, Jung-Joo
    • Journal of Life Science
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    • v.18 no.2
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    • pp.264-272
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    • 2008
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Acanthogobius hasta were characterized by biochemical, immunochemical and kinetic methods. The activities of LDH in skeletal muscle and eye tissues were 65.30 and 53.25 units, but LDH activities in heart and liver tissues were very low. LDH/CS (EC 4.1.3.7, citrate synthase) in skeletal muscle was the highest as 22.29. Specific activities of LDH in brain, eye and skeletal muscle were 56.45, 38.04 and 11.0 units/mg, respectively. The LDH isozymes in tissues were separated by polyacrylamide gel electrophoresis after immunoprecipitation with antiserum against $A_4,\;B_4$ eye-specific $C_4$ and liver-specific $C_4$. LDH $AC_4$ isozymes were detected predominantly in skeletal muscle, brain and eye tissues, and $B_4$ isozyme was detected in heart. Anodal eye-specific $C_4$ and cathodal liver-specific $C_4$ were coexpressed in A. hasta. The eye-specific $C_4$ isozyme showed higher activity in eye tissue, but liver-specific $C_4$ isozyme showed lower activity in liver. As a result, one part of molecular structures in $A_4\;and\;C_4,\;A_4\;and\;B_4$, and eye-specific $C_4$ and liver-specific $C_4$ were similar, but in $B_4\;and\;C_4$ were different with each other. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The LDH $A_4$ isozyme of skeletal muscle was purified in the fraction from elution with NAD+ containing buffer of affinity chromatography and eye-specific $C_4$ isozyme was eluted right after $A_4$, so the structure of eye-specific $C_4$ isozyme is similar to $A_4$. And LDH activity remained 35.22-43.47% as a result of the inhibition by pyruvate, the Michaelis-Menten constant values for pyruvate was 0.080-0.098 mM, and Vmax were 153.85 units, 35.09 units in skeletal muscle and eye, respectively. Also the $B_4$ isozyme was the thermo-stablest and $C_4$ was stabler than $A_4$ isozyme. The optimum pH of LDH was 6.5. The results mentioned above indicate that isozymes in tissues showed the properties between LDH $A_4\;and\;B_4$ isozyme as A. hasta was adapted to hypoxic conditions. Also LDH seems to function more effectively under anaerobic condition because LDH in skeletal muscle and eye tissues have high affinity for pyruvate.