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Qualitative and quantitative PCR detection of insect-resistant genetically modified rice Agb0101 developed in korea (해충저항성 유전자변형 벼 Agb0101에 대한 PCR 검정)

  • Shin, Kong-Sik;Lee, Jin-Hyoung;Lim, Myung-Ho;Woo, Hee-Jong;Qin, Yang;Suh, Seok-Cheol;Kweon, Soon-Jong;Cho, Hyun-Suk
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.18-26
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    • 2013
  • Genetically modified (GM) rice Agb0101, which expresses the insecticidal toxin modified cry1Ac (mcry1Ac1) gene, was developed by the Rural Development Administration in Korea. To monitor the probable release of Agb0101 in the future, it is necessary to develop a reliable detection method. Here, we developed the PCR detection method for monitoring and tracing of GM rice. The primer pair (RBEgh-1/-2) from a starch branching enzyme (RBE4) gene was designed as an endogenous reference, giving rise to an expected PCR amplicon of 101 bp. For the qualitative PCR detection, construct- and event-specific primers were designed on the basis of integration sequence of T-DNA. Event-specific PCRs amplified specifically 5'- or 3'-junction region spanning the native genome DNA and the integrated gene construct, while none of amplified product was shown on crops, rice varieties, and other insect-resistant transgenic rice lines. The event-specific real-time PCR method was performed using TaqMan probe and plasmid pRBECrR containing both rice endogenous gene RBE4 sequence and 5'-junction sequence as the reference molecule. The absolute limit of quantification (LOQ) of real-time PCR was established with around 10 copies for one plasmid molecule pRBECrR. Thereafter, the different amounts of transgenic rice (1, 3, 5, and 10%, respectively) were quantified by using the established real-time PCR method, with a range below 19.55% of the accuracy expressed as bias, 0.06-0.40 of standard deviation (SD) and 3.80-7.01% of relative standard deviations (RSD), respectively. These results indicate that the qualitative and quantitative PCR methods could be used effectively to detect the event Agb0101 in monitoring and traceability.

Genetic Analysis of Some Polymorphic Isozymes in Pinus densiflora (I) -Inheritance of Glutamate-Oxalate Transaminase and Leucine Aminopeptidase, and Linkage Relationship among Allozyme Loci- (소나무의 몇가지 다형적(多形的) 동위효소(同位酵素)의 유전분석(遺傳分析)(I) -Glutamate-Oxalate Transaminase 와 Leucine Aminopeptidase의 유전(遺傳)과 동위효소(同位酵素) 유전자좌(遺傳子座) 간(間)의 연관관계(連關關係)-)

  • Kim, Zin Suh;Hong, Yong Pyo
    • Journal of Korean Society of Forest Science
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    • v.58 no.1
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    • pp.1-7
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    • 1982
  • Megagametophyte and embryo tissue of Pinus densiflora were subjected to study the inheritance of glutamate-oxalate transaminase(GOT) and leucine aminopeptidase(LAP), and linkage relationship among isozyme loci coding both enzymes by starch gel zone-electrophoresis. Four zones of activity were observed for GOT. No variation was found in the fastest migrating zone (GOT-A). Electrophoretic phenotypes of the other two zones (GOT-B and GOT-C) showed 1:1 segregation ration, suggesting that each zone is controlled by a single locus. Foru and three alleles were identified at both loci respectively. The isozyme pattern of the fourth zone(GOT-D), migrated cathodally, coincided precisely with that of GOT-C. Whether the two zones are controlled by the same locus or by two tightly linked loci remained unknown. In all three variant GOT zones, heterozygoes embryos produced triple band patterns, indicating that GOT isozyme in Pinus densiflora is a dimer. Two zones of activity stained for LAP were found. The segregation of the two zones (LAP-A and LAP-B) suggested that tow loci control each of both isozymes. Two and three alleles were identified at both loci. GOT-B and LAP-B were found to be tightly linked, showing an average recombination frequency of 12.5 percent. Slight deviation from independent assortment was observed between GOT-B and GOT-C, with recombination frequency of 41 percent.

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Developmental Capacity of Bovine Follicular Oocytes after Ultra-Rapid Freezing by electron Microscope Grid II.Cryopreservation of In Vitro Matured Bovine Oocytes (Electron Microscopic Grid를 이용한 초급속 동결이 소 난포란의 발달능에 미치는 영향. II. 체외 성숙된 소 미수정란의 동결에 관한 연구)

  • 김은영;김남형;이봉경;윤산현;박세필;정길생;임진호
    • Korean Journal of Animal Reproduction
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    • v.22 no.1
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    • pp.1-9
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    • 1998
  • This study was carried out to confirm whether the developmental capacity of bovine mature oocytes frozen ultra-rapidly using electron microscopic(EM) grids and EFS30 can be obtained, and whether the cryoprotectants and the freezing method used in this study effect detrimentally to the bovine oocytes by indirect immunocytochemistry. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. The results obtained in this experiment were summarized as follows: When the effects of cryoprotectant and freezing procedure on the microtuble, micrfilament and chromatin morphology of oocytes were evaluated using indirect immunocytochemistry, the results of freezing as well as exposure group were not different with that of the control oocytes. When the fertilization abnormality after ultrarapid freezing of bovine mature oocytes was examined by Hoechst staining, the rates of total penetration(96.7, 9.0%), normal two pronuclei formation(74.6, 68.9%) and mean number of sperm / oocyte(1.50, 1.44) were not different between control and freezing group. In addition, when the developmental capacity of frozen-thawed of oocytes(85.5%) was survived, 74.5% of them were cleaved and 31.4% of cleaved embryos were developed to blastocyst. These data were similar to those of the control(76.0%, 34.6%) and exposure(74.5%, 33.0%) except survival rates. Also, when the total cell number of blastocysts produced from the each treatment at day after IVF was examined by hoechst staining, there were not different among groups. There results demonstrate that developmental capacity of frozen-thawed bovine mature oocytes can be successfully obtained by ultra-rapid freezing method using EM grid and EFS30 solution.

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Differences in Physicochemical and Textural Properties of Germinated Brown Rice in Various Rice Varieties (발아현미의 이화학적 특성 및 취반 특성)

  • Cho, Dong-Hwa;Park, Hye-Young;Lee, Seuk-Ki;Park, Jiyoung;Choi, Hye-Sun;Woo, Koan-Sik;Kim, Hyun-Joo;Sim, Eun-Yeong;Won, Yong-Jae;Lee, Dong-Hyun;Oh, Sea-Kwan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.62 no.3
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    • pp.172-183
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    • 2017
  • Germinated brown rice (GBR) has received considerable attention over the last decade as a means of enhancing the nutritional value and health-promoting functions of rice. The effect of germination on the physicochemical and textural properties of brown rice (BR) was investigated in different rice varieties (Samkwang, Misomi, Chindeul, and Hyeonpum). Cooking properties, such as water absorption, expanded volume, and soluble solids were significantly increased by germination in all rice varieties. Textural properties (hardness, toughness, adhesiveness, and stickiness) of cooked samples were determined using a texture analyzer. Hardness and toughness were decreased by germination, whereas stickiness and adhesiveness increased significantly. These results revealed that germination leads to improvements in the cooking and eating properties of BR. In Rapid Visco Analyzer (RVA), significant reductions were observed for peak viscosity, break down, set back, and final viscosity after germination. Although the amylose content of Misomi was slightly decreased by germination, that of other varieties increased significantly. Germination induced no noticeable change in the average chain length of amylopectin in Misomi, Chindeul and Hyeonpum, but led to a significant increase in Samkwang.

Effects of Saponin Contained Plant Extracts on Ruminal Fermentation Characteristics and Methane Production (Saponin 함유 식물 추출물의 첨가가 반추위 발효성상과 메탄생성에 미치는 영향)

  • Ok, Ji-Un;Baek, Youl-Chang;Kim, Kyoung-Hoon;Lee, Sang-Cheol;Seol, Yong-Joo;Lee, Kang-Yeon;Choi, Chang-Weon;Jeon, Che-Ok;Lee, Sang-Suk;Lee, Sung-Sil;Oh, Young-Kyoon
    • Journal of Animal Science and Technology
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    • v.53 no.2
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    • pp.147-154
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    • 2011
  • This study was conducted to evaluate the effects of saponin contained plant extracts on in vitro rumen fermentation characteristics and methane production. Ruminal fluid was collected from rumen cannulated Hanwoo steers fed rice straw and concentrate (5:5). Collected rumen fluids, corn starch and buffer including saponin contained plant extracts (ginseng, Ogapi, soapwort, tea plant and yucca; 0.5%/15 ml) were incubated at $39^{\circ}C$ for 24 h. All incubations were repeated five times. Rumen pH in all treatments was lower (p<0.05) compared with that of the control (no addition) during incubation time. The concentration of total VFA in all treatments was higher (p<0.05) than that of the control after 12h incubation. Compared with the control, the concentration of acetate and propionate in all treatments was lower and higher after 6h incubation, respectively. The concentration of $NH_3$-N in all treatments was lower (p<0.05) than that of the control except for Ogapi or yucca extracts supplementation. The number of protozoa in all treatments was significantly (p<0.05) lower than that of the control except for soapwort extract supplementation. The total gas production and methane production in all treatments was higher (p<0.05) and lower (p<0.05) compared with the control, except for ogapi or soapwort extracts supplementation after 12h incubation, respectively. Therefore, reduction in methane production by saponins may could be results from decreased protozoal population without any negative in vitro fermentation.

Mucilage Separation of Korean Yam Using Microparticulation/Air Classification Process (초미세분쇄/공기분급을 이용한 마의 점질물 분리)

  • Lee, Boo-Yong;Park, Dong-June;Ku, Kyung-Hyung;Kim, Hyun-Ku;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.26 no.5
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    • pp.596-602
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    • 1994
  • To separate and concentrate mucilage from yam(Dioscorea batatas DECNE), yam was dried, microparticulated using impact mill and air-classified at different air classifying wheel speed(ACWS) in classifier. As ACWS increased from 5,000 rpm to 22,500 rpm, the contents of dietary fiber, protein and lipid of air classified microparticles(ACM) increased remarkably. Especially the ACM with ACWS over 15,000 rpm showed 36.41% dietary fiber and 16.66% protein. The dietary fiber and protein components were concentrated to $2.5{\sim}9.0$ times as compared with whole yam powder. Concomitantly the non-fibrous carbohydrate decreased from 88.31% to 16.84. The damaged starch(%), WSI and WAI of ACM of ACWS over 15,000 rpm were $1.5{\sim}3.0$ times higher than those of ACM under ACWS 15,000 rpm. The apparent viscosity of ACM was 0.0800 Pa s over ACWS 15,000 rpm and 0.0080 Pa s under ACWS 15,000 rpm. Judging from viscosity of ACM, the mucilage component of yam was concentrated to 10 times. In conclusion, the optimum process to separate and concentrate the mucilage from yam consisted of the microparticulation to $5{\sim}30{\mu}m$ and the air-classification at ACWS over 15,000 rpm.

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The Additional Effects of Various Materials on Microwave Heating Property of Frozen Dough (품질개량제 첨가가 냉동반죽의 Microwave 가열특성에 미치는 영향)

  • Kim, Eun-Mi;Han, Hye-Kyung;Kim, In-Ho
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.873-881
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    • 2005
  • This study was conducted to improve the properties of frozen dough foods (buns and noodles etc.) on the quality deterioration with microwave oven cooking. Microwave is a useful cooking method, but it quickly takes moisture from food surface and makes lowering food quality abruptly. For improvement of these problems, mixing doughs with addition of various additives of 34 types manufactured respectively; starches, modified starches, gums and emulsifiers etc. Each mixing dough produced in sheet type $(30{\times}30{\times}1mm)$ and steamed them, was quickly froze at $-70^{\circ}C$ and packed with polyethylene. Packed samples kept at $-20^{\circ}C$ for 48 hours. After they were steam or microwave treatment packed or non-packed with polyethylene, studied for improvement effects of quality as sensory evaluation and selected 6 type additives; modified starches (TA, ST), gums (AR, GA) and emulsifiers (E, S1) as improvement agent. Because moisture loss from microwave oven cooking leads to quality deterioration of frozen dough foods, additive, such as including starches, modified starch, gums, and emusifiers were added to improve dough properties. Amylogram, scanning electron microscopy, textural analysis, and differential scanning calorimetry revealed addition of additives improved textural properties including surface-hardening of frozen dough foods compared to the control.

THE CHANGE OF THE CONFIGURATION OF HYDROXYAPATITE CRYSTALS IN ENAMEL BY CHANGES OF PH AND DEGREE OF SATURATION OF LACTIC ACID BUFFER SOLUTION (유산 완충용액의 pH 및 포화도 변화에 따른 법랑질 내 수산화인회석 결정 형태의 변화)

  • Chon, Young-Eui;Jung, Il-Young;Roh, Bung-Duk;Lee, Chan-Young
    • Restorative Dentistry and Endodontics
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    • v.32 no.6
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    • pp.498-513
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    • 2007
  • Since it was reported that incipient enamel caries can be recovered, previous studies have quantitatively evaluated that enamel artificial caries have been, remineralized with fluoride showing simultaneously the increase of width of surface layer and the decrease of width of the body of legion. There is, however, little report which showed that remineralization could occur without fluoride. In addition, the observations on the change of hydroxyapatite crystals also have been scarcely seen. In this study, enamel caries in intact premolars or molars was induced by using lactic acidulated buffering solutions over 2 days. Then decalcified specimens were remineralized by seven groups of solutions using different degree of saturation(0.212, 0.239, 0.301, 0.355) and different pH(5.0, 5.5, 6.0) over 10 days. A qualitative comparison to changes of hydroxyapatite crystals after fracturing teeth was made under SEM(scanning electron microscopy) and AFM(atomic force microscopy). The results were as follows: 1. The size of hydroxyapatite crystals in demineralized area was smaller than the normal ones. While the space among crystals was expanded, it was observed that crystals are arranged irregularly. 2. In remineralized enamel area, the enlarged crystals with various shape were observed when the crystals were fused and new small crystals in intercrystalline spaces were deposited. 3. Group 3 and 4 with higher degree of saturation at same pH showed the formation of large clusters by aggregation of small crystals from the surface layer to the lesion body than group 1 and 2 with relatively low degree of saturation at same pH did. Especially group 4 showed complete remineralization to the body of lesions. Group 5 and 6 with lower pH at similar degree of saturation showed remineralization to the body of lesions while group 7 didn't show it. Unlike in Group 3 and 4, Group 5 and 6 showed that each particle was densely distributed with clear appearance rather than crystals form clusters together.

Rice Fermentation by Korean Amylolytic Bifidobacterium spp. (한국인유래의 Amylolytic Bifidobacterium 에 의한 쌀발효)

  • Park, Jong-Hyun;Song, Hey-Kyung;Ahn, Jun-Bae;Ji, Geun-Eok;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.581-587
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    • 1997
  • For bifidus fermentation food, gelatinized rice solution was fermented without liquefaction/saccharification by amylolytic Bifidobacterium spp. isolated from Korean. Eighteen amylolytic Bifidobcterium on the starch agar were isolated from 38 Korean and four strains were finally selected as good amylase producers. The most enzyme-producing strain of Bif. sp. FBD-12 secreted extracellular amylase of 0.17 U/mg and intracelluar amylase of 1.8 U/mg. Three strains of Bif. sp. FBD-12, Bif. sp. FBD-16 and Bif. sp. FBD-17 also showed good growth on pH controlled media by HCI/acetic acid to pH 5.0 while Bif. sp. FBD-6 was not so tolerant that viable cell counts reduced to $10^2\;CFU/mL$ times on the media. Initial cell number of $10^6\;CFU/mL$ for those strains reached to $10^9\;CFU/mL$ on the rice medium supplemented with yeast extract (0.2%) and cysteine (0.05%). Ascorbic acid instead of cysteine was added to the medium for improving off-flavour and the best growth was shown at 0.1% addition. Isolated soybean proteins (ISP) of 3% accelerated the growth of the strains. Maximum count of $10^9\;CFU/mL$ reached within 12 hour fermentation on the rice medium with ascorbic acid and isolated soybean protein instead of 32 hours on the cysteine medium, and total acidity increased from 0.5% to 1% on each media. Reducing sugar in the ascorbic acid/ISP cultures generally increased especially 2 mg/mL to 15.5 mg/mL for Bif. sp. FBD-6. From sensory evaluation, the products showed good acceptability so that it suggested possibility of development of bifidus-fermented rice food.

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Changes in the Physicochemical Characteristics of Green Wheat during Maturation (성숙과정 중 녹색밀의 이화학적 특성변화)

  • Kim, Min-Chul;Lee, Ka-Soon;Lee, Byung-Jin;Kwon, Byung-Gu;Ju, Jung-Il;Gu, Ja-Hyeong;Oh, Man-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1307-1313
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    • 2007
  • In order to evaluate the utilization of immature green wheat kernels as food processing material, these experiments were carried out to analyze general composition, free sugars, free amino acids, mineral content and color of 3 wheat varieties according to their heading dates. Contents such as moisture, crude protein, crude lipid and ash were gradually decreased according to the days after heading of 3 wheat varieties, while crude fiber and starch were gradually increased. Maltose and maltotriose contents in immature green wheat kernels were high but its rapidity decreased by closing to ripeness. Contents of free amino acids showed a tendency to decrease, among which alanine content was the highest around 25 days after heading, and then glutamic acid, GABA, glycine in order. Contents of total free amino acid were decreased by closing to ripeness. Among 3 wheat varieties, its contents of Guru wheat was the highest. As wheat matured, ${\beta}-glucan$ contents were decreased from 1.5% to 0.28% on 43 days after heading. Mineral contents of 3 wheat varieties were generally increased by closing to ripeness after coming into ears, and K, Mg were especially prominent. As wheat matured, lightness was decreased, while redness and yellowness were increased. Considering chemical composition and color, the immature green wheat were produced through blanching the spikes harvested before the yellow ripe stage. The harvested green wheat cereals is able to eat raw wheat or cook it as food processing material.