• Journal of Plant Biotechnology
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• v.30 no.3
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• pp.233-239
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• 2003

We have developed a reliable and high-frequency genetic transformation and regeneration system via somatic embryogensis of Eleutherococcus sessiliflorus. Embryogenic callus obtained from seed were co- cultivated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring genes for intron-$\beta$-glucoronidase(GUS), kanamycin and hygromycin resistance. Following co-cultivation, two types of samples(fine embrogenic calli and early globular embryo clusters) were cultivated on Murashige and Skoog(MS) medium containing 1 mg/L2.4-D for 3day in dark. Transient expression of GUS gene was found to be higher in the early globular embryo clusters than in the embryogenic calli. Also, co-cultivated period affected expression of GUS gene; the best result was obtained when globular embryo clusters were co-cultivated with Agrobacterium for 3 days. Subsequently, this callus transferred to selective MS medium containing 1mg/L2.4-D, 50mg/L kanamycin or/and 30mg/L hygromycin and 300mg/L cefortaxime. These embryogenic calls were subcultured to the same selection medium at every 2 weeks intervals. Approximately 24.5% of the early globular embryos co-cultivated with Agrobacterium for 3days produced kanamycin or/and hygromycin-resistant calli. Transgenic somatic embryos were converted into plantlets in half strength MS medium supplemented with 3mg/L GA$_3$ kanamycin and were confirmed by GUS histochemical assay and polymerase chain reaction analysis. Genomic Southem blot hybridization confirmed the incorporation of NPT II gene into the host genome.

• Journal of Microbiology and Biotechnology
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• v.9 no.5
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• pp.687-690
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• 1999

Bacillus stearothermophilus No. 236 isolated from the soil is a strong xylan degrader producing all the xylanolytic enzymes. However, the strain was discovered to be highly intractable to its transformation. In the present study, we have developed a reliable method for transformation of B. stearothermophilus No. 236 by a systematic examination of several factors which might have an influence on the efficiency of electrotransformation. Notably, we found that the most critical factor influencing the transformation efficiency (TE) was the incubation temperature after pulsing, with its optimum incubation of $37^{\circ}C.\; At\; 50^{\circ}C$, the optimum growth temperature of the B. stearothermophilus strain, the transformants could not be obtained at a recognizable level. The combination of field strength of 7.5 kV/cm along with pulse duration of 10 msec (resistance of $400{\Omega}\; and\; capacitance\; of\; 25{\mu}F$) was shown to be the best electrical parameters at the incubation temperature of $37^{\circ}$. A higher TE was obtained when the cells were harvested at an early-exponential phase. Twenty percent of PEG-8000 in a suspension buffer and an addition of 0.1% glycine in the growth medium resulted in about 4-fold and 3-fold increases in TE, respectively. We also found that the plasmid DNA which had been cycled through the host B. stearothermophilus cells enhanced TE by one order of magnitude higher. Under the presently described conditions, $2.5{\times}10^{5} transformants per ${\mu}g$ DNA was attained.

• International Journal of Oral Biology
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• v.39 no.4
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• pp.207-213
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• 2014

Antimicrobial actions of reactive oxygen/nitrogen species (ROS/RNS) derived from products of NADPH oxidase and inducible nitric oxide (NO) synthase in host phagocytes inactivate various bacterial macromolecules. To cope with these cytotoxic radicals, pathogenic bacteria have evolved to conserve systems necessary for detoxifying ROS/RNS and repairing damages caused by their actions. In response to these stresses, bacteria also induce expression of molecular chaperones to aid in ameliorating protein misfolding. In this study, we explored the function of a newly identified chaperone Spy, that is localized exclusively in the periplasm when bacteria exposed to conditions causing spheroplast formation, in the resistance of Salmonella Typhimurium to ROS/RNS. A spy deletion mutant was constructed in S. Typhimurium by a PCR-mediated method of one-step gene inactivation with ${\lambda}$ Red recombinase, and subjected to ROS/RNS stresses. The spy mutant Salmonella showed a modest decrease in growth rate in NO-producing cultures, and no detectable difference of growth rate in $H_2O_2$ containing cultures, compared with that of wild type Salmonella. Quantitative RT-PCR analysis showed that spy mRNA levels were similar regardless of both stresses, but were increased considerably in Salmonella mutants lacking the flavohemoglobin Hmp, which are incapable of NO detoxification, and lacking an alternative sigma factor RpoS, conferring hypersusceptibility to $H_2O_2$. Results demonstrate that Spy expression can be induced under extreme conditions of both stresses, and suggest that the protein may have supportive roles in maintaining proteostasis in the periplasm where various chaperones may act in concert with Spy, thereby protecting bacteria against toxicities of ROS/RNS.

• Preventive Nutrition and Food Science
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• v.7 no.2
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• pp.168-173
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• 2002

Gastrointestinal infections kill over two million people each year, and pathogen contamination of livestock causes many cases of food poisoning. Two candidate intestinal probiotic strains, L. rhamnosus GR-1 and L. fermentum RC-14 were found to inhibit the growth of Salmonella typhi, Shigella dysenteriae, E. coli O157:H7, Listeria monocytogenes, L. innocua, Enterococcus faecalis, and Bacteroides fragilis. In a series of mouse experiments, L. rhamosus GR-1 and L rhamnosus GG protected against S. typhimurium infection and translocation to the liver and spleen, reduced mortality and induced intestinal phagocytic and immunoglobulin responses. In a second series of experiments, the combination of L. rhamnosus GR-1 and L. fermentum RC-14 was superior to L. rhamnosus GG and placebo in protecting the mice from the lethal effect of salmonella. In summary, the use of combinations of probiotic lactobacilli as dietary supplements or foods could be considered for people at high risk of salmonella intestinal infection. Given the post-infection complications that can arise, such natural methods warrant further exploration especially given the increasing problem of antibiotic resistance and the lack of alternative measures available to many developing countries.

• Journal of Microbiology and Biotechnology
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• v.28 no.10
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• pp.1736-1748
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• 2018

Brucella abortus can survive and replicate within host macrophages, and great efforts have been made to demonstrate the genes involved in pathogenicity, such as internalization, in Brucella research. Here, intracellular responses were compared between THP-1 macrophage cells stimulated with B. abortus wild-type and four mutants (C1, C10, C27, and C32) using microarray to demonstrate the role of genes related to intracellular survival and replication. These mutants were generated by deleting genes encoding BAB_RS13225 (4-hydrobenzoate 3-monooxygenase, PHBH), BAB_RS00455 (heme exporter protein cytochrome C, CcmC), BAB_RS03675 (exopolyphosphatase, PPX), and BAB_RS13225 (peptidase M24). The results showed that mutants C1 and C10 induced significant suppression of survival levels and cytokine expression relative to wild-type in the THP-1 macrophage cells. These findings suggest that the BAB_RS13225 and BAB_RS00455 genes play important roles in survival within human macrophages. Conversely, mutants C27 and C32 induced significantly higher survival level than wild-type in the cells inhibiting cellular signal transduction. It is assumed that the BAB_RS03675 and BAB_RS13225 genes play a role in cellular resistance to B. abortus. Therefore, the disrupted genes are involved in B. abortus intracellular growth, and especially in its survival, and they could be effective targets for understanding the intracellular bacterium, B. abortus.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.57 no.2
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• pp.113-126
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• 2012

Northern corn leaf blight caused by Exserohilum turcicum Pass is considered the most important disease infecting corn (Zea mays L.) in the Peoples' Republic of Korea (North Korea). It contributes to the food shortage in North Korea. The objectives of the current research were to study resistance expression and responses of corn crosses made between ten hybrids from North Korea and inbreeding lines ($S_{3-4}$ stage) from the Republic of Korea (South Korea). The experiments were conducted in six trials with a total of 184 crosses including two commercial hybrids in each trial. The trials were conducted at two locations in North Korea (Mirim and Eunsan) and one location in South Korea (Gunwi) under natural infestation of E. turcicum. Host plant responses were rated on a scale of 1 (highly tolerant) to 9 (highly susceptible). A total of 111 crosses (62.4%) showed significant tolerant or susceptible response variations among three locations; 42 crosses (22.8%) at two locations and 69 crosses (39.0%) at one location, respectively. At least 8 crosses of high level of tolerance and 12 crosses of high level of susceptibility showed significantly different biotic responses (P = 0.05). The results of the current study and historical reviews of E. turcicum epidemics in both North and South Korea suggest that breeding of tolerance with quantitatively inherited genes should be carried out for a sustainable corn production in North Korea.

• Journal of Life Science
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• v.14 no.2
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• pp.209-214
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• 2004

Cytotoxic anticancer chemotherapeutic agents generally produce severe side effects, while reducing host resistance to cancer and infections, especially through the destruction of lymphoid and bone marrow cells. In this study, we have investigated the effect of chitosan on cytotoxic activity against cancer cells and life span in mice. The direct cytotoxicity of chitosan or chitosan-combinated chemotherapeutic agents for tumor cells was observed. In addition, the effect of life span extention was counted on sarcoma 180 mice injected with chitosan-combinated mitomycin C. The effect of growth inhibion for cancer cells, K562 and Yac-1 was shown in the cytotoxicity test of chitosan or chitosan-combinated chemotherapeutic agents. Also, the effect of life span extension was observed on sarcoma 180 mice injected with chitosan-combinated mitomycin C. Our results suggest that life span extension in sarcom 180 mice exposed with chitosan-combinated chemotherapeutic agents showed the probability of its usefulness for cancer therapy if more research results were accumulated.

• Journal of the Korean Home Economics Association
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• v.30 no.3
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• pp.91-99
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• 1992

Cell structure is based on proteins. Since the antibody is proteous substance, the continous low protein feeding decreases the resistance of host against pathogenic agents. The present study was designed to investigate the infectivity of protozoa to rats which were fed with variously prescribed diets. Experimental group was divided into 4 groups according to the level of casein in the diet, group I: casein 0%, group II: casein 5%, group III: casein 15%, group IV: casein 30%. Each animal was fed for 5 weeks followed by inoculation of protozoa in cecum and sacrified each 1 week later of the infection. Each diet group, non infected with protozoa was recognized as the control. Result are summerized as follows : 1. All the rats of group I died in 2∼4 weeks and 2 of 12 rats in group II were also died in the period. 2. The growth rate and FER were high in group III and IV compared with group II. Therefore low protein feeding decrease growth and feed efficaly ratio(FER). 3. The pH of caecal contents between the infected group and control showed no difference, but the values of group III and IV were higher than the group II. Low pH of the caecal contents provides a suitable condition for determining their susceptibility to Entameoeba histolytical trophozoite. 4. Amounts of serum total protein in group II, III and IV showed no significant difference with the control and infected group, but amounts in group III and IV were higher than the group II. Therefore, continuous low protein feeding decrease serum total protein. 5. Albumin, ${\alpha}$1, gloulin, ${\alpha}$2 globulin, ${\beta}$ globulin, ${\gamma}$ gloulin of group III and IV were all high to compare than the group II. Albumins of group III and IV of control was higher than infected group, but there was no difference in ${\gamma}$ globulin between the infected and control group.

• Korean Journal of Poultry Science
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• v.34 no.4
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• pp.301-318
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• 2007

Marek's disease (MD) is a highly contagious lymphoproliferative disease of poultry caused by the oncogenic herpesvirus designated Marek's disease virus (MDV). MD has a worldwide distribution and is thought to cause an annual loss over US$ one billion to the poultry industry. Originally described as a paralytic disease, today MD is mostly manifested as an acute disease with tumors in multiple visceral organs. MD is controlled essentially by the widespread use of live vaccines administered either in ovo into 18-day-old embryos or into chicks immediately after they hatch. In spite of the success of the vaccines in reducing the losses from the disease in the last 30 years, MDV strains have shown continuous evolution in virulence acquiring the ability to overcome the immune responses induced by the vaccines. During this period, different generations of MD vaccines have been introduced to protect birds from the increasingly virulent MDV strains. However, the virus will be countered each new vaccine strategy with ever more virulent strains. In spite of this concern, currently field problem from MD is likely to be controled by strategy of using bivalent vaccine. But, potential risk factors for outbreak of MD are still remained in this condition. The major factors can be thought that improper handling and incorrect administration of the vaccine, infection prior to establishment of immunity, suppression of immune system by environmental stress and outbreaks of more virulent MDV strain by using vaccine and genetic resistance of host.

• Food Science of Animal Resources
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• v.40 no.5
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• pp.746-757
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• 2020

Enterotoxigenic Escherichia coli (ETEC) is the major pathogenic E. coli that causes diarrhea and edema in post-weaning piglets. In this study, we describe the morphology and characteristics of ØCJ19, a bacteriophage that infects ETEC, and performed genetic analysis. Phage ØCJ19 belongs to the family Myoviridae. One-step growth curve showed a latent phase of 5 min and burst size of approximately 20 phage particles/infected cell. Phage infectivity was stable for 2 h between 4℃ and 55℃, and the phage was stable between pH 3 and 11. Genetic analysis revealed that phage ØCJ19 has a total of 49,567 bases and 79 open reading frames (ORFs). The full genomic sequence of phage ØCJ19 showed the most similarity to an Escherichia phage, vB_EcoS_ESCO41. There were no genes encoding lysogeny, toxins, virulence factors, or antibiotic resistance in this phage, suggesting that this phage can be used safely as a biological agent to control ETEC. Comparative genomic analysis in terms of the tail fiber proteins could provide genetic insight into host recognition and the relationship with other coliphages. These results showed the possibility to improve food safety by applying phage ØCJ19 to foods of animal origin contaminated with ETEC and suggests that it could be the basis for establishing a safety management system in the animal husbandry.