• Title/Summary/Keyword: Toxin gene

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Molecular characterization of lepidopteran pest-resistant transgenic rice events expressing synthetic Cry1Ac

  • Lee, Kyeong-Ryeol;Shin, Kong Sik;Suh, Seok Cheol;Kim, Ki Young;Jeon, Yong Hee;Park, Beom Seok;Kim, Ju-Kon;Kweon, Soon-Jong;Lee, Yeon-Hee
    • Plant Biotechnology Reports
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    • v.3 no.4
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    • pp.317-324
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    • 2009
  • The insecticidal toxin gene of Bacillus thuringiensis (Bt) is one of the most commonly used in the development of genetically modified (GM) crops. In this research, we analyzed Bt rice showing lepidopteran pest-resistance. The Bt gene is a synthetic Cry1Ac composed of optimal codons for plants, and the Bt protein is targeted to the chloroplast by a transit peptide. Three Cry1Ac rice events (C103-3, C127-1, and C7-1) were analyzed for molecular characterization. C103-3 contains two copies of T-DNA where the left border (LB) region is truncated. Both C7-1 and C127-1 have a single copy of T-DNA, but a part of the vector backbone DNA is inserted into the genome of C127-1; thus, only C7-1 had intact T-DNA. Progenies of C7-1 crossed with the original cultivar, Nakdong, and double-haploid lines from anther culture of lines crossed with the elite cultivar, Dongjin, were analyzed for T-DNA flanking genomic DNA and genotyping. Results showed that an intact T-DNA region without the vector backbone was inserted into the genome and was stably inherited through generations. The C7-1 homozygous event could be used as breeding material to develop GM rice with pest resistance.

Profiles of Toxin Genes and Antibiotic Susceptibility of Bacillus cereus Isolated from Perilla Leaf and Cultivation Areas (들깻잎과 생산환경에서 분리한 Bacillus cereus의 독소 유전자와 항생제 감수성 분석)

  • Kim, Se-Ri;Lee, Ji-Young;Lee, Seo-Hyun;Ryu, Kyoung-Yul;Park, Kyeong-Hun;Kim, Byung-Seok;Yoon, Yo-Han;Shim, Won-Bo;Kim, Kyoung-Yul;Ha, Sang-Do;Yun, Jong-Chul;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.134-141
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    • 2011
  • Two-hundred Bacillus cereus isolated from perilla leaf cultivation areas in Miryang, Korea were investigated for toxin genes and antibiotic susceptibility. Toxigenic patterns of isolates were identified to be 11 groups through toxin gene profiles. 21% of strains isolated from the perilla leaves had both enterotoxin and emetic toxin. Toxin genes entFM (100%), nheA (100%) and hblA, C, D (65.5%) were frequently found in the perilla leaves, whereas EM (21.0%) was less common. Most isolates were susceptible to 10 antibiotics, but they were highly resistant to penicillin (100%), ampicillin (100%), oxacillin (94.9%), amoxicillin-clavulanic acid (95.6%), cefazolin (78.2%), and rifampicin (58.0%). These results indicate that food-borne outbreak caused by B. cereus might lead to diarrhea and emetic syndromes.

Effects of Helicobacter pylori Antigen on Producton of Transforming growth factor-$\beta$1 and Nitric oxide in Human Fibroblast (사람성유아세포의 Transforming growth factor-$\beta$1과 Nitric oxide 생성에 미치는 Helicobacter pylori 항원의 효과)

  • 박무인;박선자;구자영;김광혁
    • Journal of Life Science
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    • v.11 no.2
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    • pp.181-189
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    • 2001
  • Cytokines are hormone-like proteins which mediate and regulast inflammatory and immune responses. Transforming growth factor -$\beta$1(TGF-$\beta$) plays an important role in the control of the immune response and wound healing, and in the development o various tissues and organs, Nitric oxide(NO) is major messenger molecule regulating immune function and blood vessel dilation and serving as a neurotransmitter in the brain and peripheral nervous system. Also, NO is to be a potent mutagen that cause mutation in the p53 tumor suppressor gene in early phases of human gastric carcinogenesis. The purpose of this study was to investigate the effect of Helicobacter phlori lystes, lipopolysaccharide (LPS), and Staphylococcus enterotoxin B(SEB) on production of TGF-$\beta$1 and NO by human fibroblasts. Primary cultured human fibroblasts were incubated with H. pylori lysates(Hp), LPs, SEB, Hp+LPS, Hp+SEB, Hp+LPS+SEB. Cultured supernatants that were collected at 24, 48 and 72 hr were assessed for TGF-$\beta$1 by enzyme-linked immunosorbent assay and NO production by quantification of nitrite ion. TGF-$\beta$1 production in fibroblasts exposed with Hp, LPS or SEB for 48 hrs was enhanced, but for 72 hrs inhibited. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB was lowered in comparison with single exposure of Hp in cases of 24 and 48 hrs incubation, but for 72 hrs decreased in Hp vaculoating toxin(+), increased in Hp vacuolating toxin(-). No production in fibroblasts increaed at all doses of LPS. But its production by exposure of SEB increased or decreased according to dose and incubation time. Also, NO production by Hp vacuolating toxin(+) increased at all doses, but its production by Hp vacuolating toxin(-) decreased. Its production by doble exposure such as Hp+LPS, Hp+SEB, Hp+LPS+SEB decreased in comparison with single exposure Hp Therefore, quantities pf TGB-$\beta$1 and NO released by human fibroblasts shows differences according to kinds of stimulants. Also, in care stimulated with same kinds of stimulants, its productions exhibit quantitative differences according to exposure times. These results suggest that the decreased of TGF-$\beta$1 in fibroblasts by mixed exposure with Hp producing vacuolating toxin and bacterial toxins such as LPS and SEB may effect negatively in healing of host tissue and increased of NO by infection oh H. pylori may related to the increased susceptibility for human gastric carcinogenesis.

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Prevalence and Toxin Genes of Food-Borne Pathogens Isolated from Toothbrush in Child Care Center (보육시설 유아 사용 칫솔의 식중독 미생물 분포 및 독소 유전자)

  • Kim, Jong-Seung;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.242-248
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    • 2015
  • This study was performed to investigate the microbiological contamination on toothbrushes, toothbrush caps, and tooth cleaning cups in the child care centers and to evaluate the toxin genes, toxin production ability and antibiotic resistance of food-borne pathogens. The average number of total aerobic bacteria and fungi were 5.3 log CFU and 3.2 log CFU. Coliform bacteria were detected in 41 (54.7%) of 75 toothbrushes, 13 (44.8%) of 29 toothbrush caps, and 29 (44.6%) of 65 tooth cleaning cups. Salmonella spp. was not detected in all of samples but Bacillus cereus was isolated from 1 (1.3%) of 75 toothbrushes and 2 (3.1%) of 65 tooth cleaning cups. Staphylococcus aureus was detected in 1 (1.5%) of 65 tooth cleaning cups. The nheA, nheB, nheC, hblC, hblD, hblA and entFM toxin genes were possessed in B. cereus isolated from toothbrush which also produce NHE and HBL enterotoxins. S. aureus was resistant to ampicillin and penicillin, while B. cereus was resistant to ${\beta}-lactam$ antibiotics. These results indicated that the sanitary conditions of toothbrushes and tooth cleaning cups in the child care centers should be improved promptly. The UV sterilization after drying and then storage in dried condition is required to improve the sanitary condition of toothbrushes and tooth cleaning cups in the child care center.

Insecticidal Activity and Histopathological Effects of Vip3Aa Protein from Bacillus thuringiensis on Spodoptera litura

  • Song, Feifei;Lin, Yunfeng;Chen, Chen;Shao, Ensi;Guan, Xiong;Huang, Zhipeng
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1774-1780
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    • 2016
  • Vegetative insecticidal proteins (Vips) are insecticidal proteins synthesized by Bacillus thuringiensis during the vegetative stage of growth. In this study, Vip3Aa protein, obtained by in vitro expression of the vip3Aa gene from B. thuringiensis WB5, displayed high insecticidal activity against Spodoptera litura aside from Spodoptera exigua and Helicoverpa armigera. Bioassay results showed that the toxicity of Vip3Aa protein against S. litura larvae statistically decreased along with the increase of the age of the larvae, with LC50 = 2.609 ng/cm2 for neonatal larvae, LC50 = 28.778 ng/cm2 for first instar larvae, LC50 = 70.460 ng/cm2 for second instar larvae, and LC50 = 200.627 ng/cm2 for third instar larvae. The accumulative mortality of 100% larvae appeared at 72 h for all instars of S. litura larvae, when feeding respectively with 83.22, 213.04, 341.40, and 613.20 ng/cm2 of Vip3Aa toxin to the neonatal and first to third instar larvae. The histopathological effects of Vip3Aa toxin on the midgut epithelial cells of S. litura larvae was also investigated. The TEM observations showed wide damage of the epithelial cell in the midgut of S. litura larvae fed with Vip3Aa toxin.

Growth Profile and Toxigenicity of Bacillus cereus in Ready-to-eat Food Products of Animal Origin

  • Oh, Mi-Hwa;Ham, Jun-Sang;Seol, Kuk-Hwan;Jang, Ae-Ra;Lee, Seung-Gyu;Lee, Jong-Moon;Park, Beom-Young;Kang, Eun-Sil;Kwon, Ki-Sung;Hwang, In-Gyun
    • Food Science of Animal Resources
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    • v.31 no.1
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    • pp.40-46
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    • 2011
  • The growth profile of Bacillus cereus in ready-to-eat (RTE) food products of animal origin was examined under different temperature and incubation conditions. In sandwiches and Kimbab, B. cereus did not grow or exhibited only minimal growth at 4 and $10^{\circ}C$, but it grew rapidly at ambient temperature. In sandwiches, B. cereus did not grow efficiently at $25^{\circ}C$, however, in ham, the main ingredient of sandwiches, B. cereus growth was observed at the same temperature, with bacterial levels reaching 7.94 Log CFU/g after incubation for 24 h at $25^{\circ}C$. Toxigenicity of B. cereus was observed only at temperatures above $25^{\circ}C$. In Kimbab, B. cereus produced toxin after 9 h at $30^{\circ}C$ and after 12 h at $25^{\circ}C$. Ingredients of sandwiches and Kimbab were collected from 3 different Korean food-processing companies to investigate the source of contamination by B. cereus. Among the 13 tested food items, 6 items including ham were found to be contaminated with B. cereus. Of these ingredients, B. cereus isolates from 3 items produced enterotoxins. None of these isolates harbored the emetic toxin-producing gene. The findings of the present study can be used for risk assessments of food products, including ham and cheese, contaminated with B. cereus.

Toxigenic Mycobiota of Small Grain Cereals in Korea

  • Lee, Theresa
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.33-33
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    • 2016
  • Mycotoxins are toxic secondary metabolites produced by fungi. They can be present in where agricultural-based commodities are contaminated with toxigenic fungi. These mycotoxins cause various toxicoses in human and livestock when consumed. Small grains including corn, barley, rice or wheat are frequently contaminated with mycotoxins due to infection mainly by toxigenic Fusarium species and/or under environment favorable to fungal growth. One of the most well-known Fusarium toxin groups in cereals is trichothecenes consisting of many toxic compounds. Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin, and various derivatives belong to this group. Zearalenone and fumonisin (FB) are also frequently produced by many species of the same genus. In order to monitor Korean cereals for contamination with Fusarium and other mycotoxigenic fungal species as well, barley, corn, maize, rice grains, and soybean were collected from fields at harvest or during storage for several years. The fungal colonies outgrown from the grain samples were identified based on morphological and molecular characteristics. Trichothecene chemotypes of Fusarium species or presence of FB biosynthetic gene were determined using respective diagnostic PCR to predict possible toxin production. Heavy grain contamination with fungi was detected in barley, rice and wheat. Predominant fungal genus of barley and wheat was Alternaria (up to 90%) while that of rice was Fusarium (~40%). Epicoccum also appeared frequently in barley, rice and wheat. While frequency of Fusarium species in barley and wheat was less than 20%, the genus mainly consisted of Fusarium graminearum species complex (FGSC) which known to be head blight pathogen and mycotoxin producer. Fusarium composition of rice was more diverse as FGSC, Fusarium incarnatum-equiseti species complex (FIESC), and Fusarium fujikuroi species complex (FFSC) appeared all at considerable frequencies. Prevalent fungal species of corn was FFSC (~50%), followed by FGSC (<30%). Most of FFSC isolates of corn tested appeared to be FB producer. In corn, Fusarium graminearum and DON chemotype dominate within FGSC, which was different from other cereals. Soybeans were contaminated with fungi less than other crops and Cercospora, Cladosporium, Alternaria, Fusarium etc. were detected at low frequencies (up to 14%). Other toxigenic species such as Aspergillus and Penicillium were irregularly detected at very low frequencies. Multi-year survey of small grains revealed dominant fungal species of Korea (barley, rice and wheat) is Fusarium asiaticum having NIV chemotype.

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A yeast Chromosomal Gene that Induces Defective Interfering Particles of L-A dsRNA Virus in $ski^-$ Host Cells ($ski^-$ 기주 세포에서 L-A dsRNA 바이러스의 defective interfering particle을 유도하는 효모 유전자)

  • 이현숙
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.75-79
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    • 1991
  • The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.

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Antagonism against Helicobacter Pylori and Proteolysis of Lactobacillus Helveticus CU631 and Strain Identification

  • Yoon, Y.H.;Won, B.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.7
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    • pp.1057-1065
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    • 2002
  • The antagonistic activities of 30 strains of lactobacilli against Helicobacter pylori were determined and Lactobacillus helveticus CU631 has been selected as the strain which possesses the strongest inhibitory effect in the disc diffusion assay showing inhibition zone diameter of $10{\pm}1.5mm$, whereas those of L. plantarum and L. fermentum have been shown to be $4.0{\pm}0.6mm$. H. pylori G88016 revealed the highest vacuolating toxin producing activity among the 8 strains, the inhibitory activity of L. helveticus CU631 in vacuolating toxin producing activity of H. pylori manifested in the co-culture of two strains and in the 5:5 mixture of supernatant of the two strains. Both L. helveticus CU631 and cell free culture supernatant had a strong inhibitory activities in urease and cytotoxin producing activities of H. pylori NCTC11637 and CJH12. An accelerated proteolytic activity of water soluble peptides by L. helveticus CU631 during the refrigeration storage has been manifested in the cream cheese. DNA seqences of 16S-23S ribosomal RNA spacer region showed typical pattern among the various strains of L. helveticus, which could be used in the identification of L. helveticus CU 631.

Toxigenic type of Clostridium perfringens isolated from chicken in Korea (닭에서 분리한 Clostridium perfringens의 독소형)

  • Park, Kyoung-yoon;Lee, Sang-un;Yoo, Han-sang;Yeh, Jae-gil
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.829-837
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    • 1996
  • Clostridium perfringens has been identified as a causative organism in necrotic enteritis in chicken. The bacterium has been classified into five toxigenic types (A through E) based on the pattern of the production of major lethal toxins. Seroneutralization with mice or guinea pigs usually has been used to type the organism. Of the types, types A and C of the bacterium had been recognized as the major pathogenic types in chicken. In this experiment, we isolated nine field strains of C perfringens from chicken showing necrotic enteritis in clinical symptoms and pathologic findings and identified by biochemical tests. In order to type the organism, a multiplex polymerase chain reaction (PCR) was used with primers on major lethal toxin genes instead of seroneutralization. Amplification of only a toxin gene with the PCR suggested that the disease in chicken was due to type A of C perfringens in Korea. Furthermore, the PCR method can be replaced with seroneutralization to type C perfringens in future.

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