• Title/Summary/Keyword: Agriculture Biotechnology

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Identification and Molecular Characterization of Novel cry1-Type Toxin Genes from Bacillus thuringiensis K1 Isolated in Korea

  • Li Ming Shun;Choi Jae-Young;Roh Jong-Yul;Shim Hee-Jin;Kang Joong-Nam;Kim Yang-Su;Wang Yong;Yu Zi Niu;Jin Byung-Rae;Je Yeon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.15-20
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    • 2007
  • To clone novel cry1-type genes from the Bacillus thuringiensis K1 isolate, about 2.4-kb-long PCR fragments were amplified with two primer sets of ATG1-F/N400-R and 1BeATG1-F/N400-R. Using PCR-RFLP, three novel cry1-type genes, cry1-1, cry1-7, and cry1-44, were obtained from B. thuringiensis K1 and the complete coding sequences of these novel genes were analyzed. The Cry1-1, Cry1-7, and Cry1-44 proteins showed maximum similarities of about 78.0%, 99.7%, and 91.0% with the Cry1Ha1, Cry1Be1, and Cry1Ac2 proteins, respectively. These novel cry1-type genes were expressed using a baculovirus expression vector system and their insecticidal activities were investigated. Whereas all three novel genes were toxic to Plutella xylostella larvae, only Cry1-1 showed insecticidal activity against Spodoptera exigua larvae.

Isolation, Screening and Identification of Swine Gut Microbiota with Ochratoxin A Biodegradation Ability

  • Upadhaya, Santi Devi;Song, Jae-Yong;Park, Min-Ah;Seo, Ja-Kyeom;Yang, Liu;Lee, Chan-Ho;Cho, Kyung-J.;Ha, Jong-K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.1
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    • pp.114-121
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    • 2012
  • The potential for ochratoxin A (OTA) degradation by swine intestinal microbiota was assessed in the current study. Intestinal content that was collected aseptically from swine was spiked with 100 ppb OTA and incubated for 6 and 12 h at $39^{\circ}C$. An OTA assay was conducted using the incubated samples, and it was found that 20% of the OTA toxin was detoxified, indicating the presence of microbes capable of OTA degradation. Twenty-eight bacterial species were isolated anaerobically in M 98-5 media and 45 bacterial species were isolated using nutrient broth aerobically. Screening results showed that one anaerobic bacterial isolate, named MM11, detoxified more than 75% of OTA in liquid media. Furthermore, 1.0 ppm OTA was degraded completely after 24 h incubation on a solid 'corn' substrate. The bacterium was identified by 16S rDNA sequencing as having 97% sequence similarity with Eubacterium biforme. The isolation of an OTA-degrading bacterium from the swine natural flora is of great importance for OTA biodegradation and may be a valuable potential source for OTA-degradation enzymes in industrial applications.

Determination of 11 Ginsenosides in Black Ginseng Developed from Panax ginseng by High Performance Liquid Chromatography

  • Sun, Bai-Shen;Gu, Li-Juan;Fang, Zhe-Ming;Wang, Chun-Yan;Wang, Zhen;Sung, Chang-Keun
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.561-564
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    • 2009
  • A high performance liquid chromatography (HPLC) method has been developed for determination of 11 ginsenosides in black ginseng (BG, white ginseng that is subjected to 9 cycles of $95^{\circ}C$ for 3 hr). After eluted by gradient elution of water-acetonitrile without buffer in 70 min, 11 ginsenosides in BG were identified. The proposed method provided good linearity ($R^2$>0.9995), accuracy (92.2-106.6%), and intra- and interday precision (RSD<2.6%). In addition, ginsenosides compositions in white, red, and black ginsengs were investigated using this method, respectively. Interestingly, in BG, the content of ginsenoside $Rg_3$ which does not existed in white ginseng was 7.51 mg/g, approximately 20 times than that in red ginseng.

Characterization of the Genes Involved in Induced Systemic Resistance in Cucumber Plants

  • Kim, Mi-Seong;Cho, Song-Mi;Im, Yang-Ju;Kim, Young-Cheol;Yang, Kwang-Yeol;Lee, Myung-Chul;Kim, Kwang-Sang;Cho, Baik-Ho
    • Korean Journal of Plant Resources
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    • v.20 no.2
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    • pp.216-219
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    • 2007
  • Root colonization by a rhizobacterium, Pseudomonas chlororaphis O6, elicited induced systemic resistance (ISR) in the leaves of cucumber plants against fungal and bacterial pathogens. To understand the role of unique genes during strain O6-mediated ISR, a suppressive subtractive hybridization method was undertaken and led to isolation of twenty-five distinct genes. The transcriptional levels of all the genes showed an increase much earlier under O6 treatment than in water control plants only after challenge with pathogen, while no difference detected on the plants without pathogen challenge. This suggests that O6-mediated ISR is associated with the priming phenomenon, an enhanced capacity for the rapid and effective activation of cellular defense responses after challenge inoculation.

Ginsenoside Rg1 Improves In vitro-produced Embryo Quality by Increasing Glucose Uptake in Porcine Blastocysts

  • Kim, Seung-Hun;Choi, Kwang-Hwan;Lee, Dong-Kyung;Oh, Jong-Nam;Hwang, Jae Yeon;Park, Chi-Hun;Lee, Chang-Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.8
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    • pp.1095-1101
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    • 2016
  • Ginsenoside Rg1 is a natural compound with various efficacies and functions. It has beneficial effects on aging, diabetes, and immunity, as well as antioxidant and proliferative functions. However, its effect on porcine embryo development remains unknown. We investigated the effect of ginsenoside Rg1 on the in vitro development of preimplantation porcine embryos after parthenogenetic activation in high-oxygen conditions. Ginsenoside treatment did not affect cleavage or blastocyst formation rates, but did increase the total cell number and reduced the rate of apoptosis. In addition, it had no effect on the expression of four apoptosis-related genes (Bcl-2 homologous antagonist/killer, B-cell lymphoma-extra large, Caspase 3, and tumor protein p53) or two metabolism-related genes (mechanistic target of rapamycin, carnitine palmitoyltransferase 1B), but increased the expression of Glucose transporter 1 (GLUT1), indicating that it may increase glucose uptake. In summary, treatment with the appropriate concentration of ginsenoside Rg1 ($20{\mu}g/mL$) can increase glucose uptake, thereby improving the quality of embryos grown in high-oxygen conditions.

Isolation and Characterization of Plutella xylostella Granulovirus Isolated in Korea

  • Wang Yong;Choi Jae Young;Kang Joong Nam;Kim Yang-Su;Choi Heekyu;Roh Jong Yul;Li Ming Shun;Jin Byung Rae;Im Dae Joon;Je Yeon Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.2
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    • pp.87-91
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    • 2005
  • We have first isolated and characterized a Plutella xylostella granulovirus ($P_xGV$) from dead larvae of P. xylostella in Korea. The granule of $P_xGV$ was ovoidal shape with an approximate measure of $350-400nm{\times}150-200nm$, and each granule contained one single rod-shape virion with a mean size of $150-180nm{\times}20-30nm$. The major granule protein, granulin, had a molecular weight of approximately 29kDa. Whereas the nucleotide sequence of the granulin gene was identical to that of previously reported $P_xGV$, nucleotide sequences of two of three putative p10 genes were slightly different from those of reported $P_xGV$. These results suggested that the $P_xGV$ isolated in this study was a novel isolate containing different genomic information.

Construction of a High-efficiency Shuttle Vector Containing the Minimal Replication Origin of Bacillus thuringiensis

  • Kang Joong Nam;Kim Yang-Su;Wang Yong;Choi Heekyu;Li Ming Shun;Shin Sang Chul;Jin Byung Rae;Roh Jong Yul;Choi Jae Young;Je Yeon Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.11 no.2
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    • pp.125-127
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    • 2005
  • In order to improve the transformation efficiency of the Bacillus thuringiensis (Bt)-Escherichia coli (E. coli) shuttle vector, pHT3101, we intended to minimize replication origin of Bt in pHT3101. For this, two modified shuttle vectors, pHT1K and pHT261, in which 2.9 kb of replication origin of Bt were shortened to 1 kb and 261 bp, respectively as previously reported. Whereas the pHT1K could efficiently transform Bt into the antibiotic resistant, no transformants were obtained with pHT261. Furthermore, pHT1K showed higher transformation efficiency compared to that of parent vector, pHT3101. Therefore, pHT1K might be a very useful Bt-E. coli shuttle vector carrying minimal replication origin of Bt.

Bombyx mori Nucleopolyhedrovirus Bacmid Enabling Rapid Generation of Recombinant Virus by In Vitro Transposition

  • Tao, Xue Ying;Choi, Jae Young;Kim, Yang-Su;Lee, Seok Hee;An, Saes Byeol;Pang, Ying;Kim, Jong Hoon;Kim, Woo Jin;Je, Yeon Ho
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.386-392
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    • 2015
  • A novel recombinant bacmid, bEasyBm, that enables the easy and fast generation of pure recombinant baculovirus without any purification step was constructed. In bEasyBm, attR recombination sites were introduced to facilitate the generation of a recombinant viral genome by in vitro transposition. Moreover, the extracellular RNase gene from Bacillus amyloliquefaciens, barnase, was expressed under the control of the Cotesia plutellae bracovirus early promoter to negatively select against the nonrecombinant background. The bEasyBm bacmid could only replicate in host insect cells when the barnase gene was replaced with the gene of interest by in vitro transposition. When bEasyBm was transposed with pDualBac-EGFP, the resulting recombinant virus, EasyBm-EGFP, showed high levels of EGFP expression efficiency compared with that of non-purified recombinant virus BmGOZA-EGFP, which was constructed using the bBmGOZA system. In addition, nonrecombinant backgrounds were not detected in unpurified EasyBm-EGFP stocks. Based on these results, a high-throughput system for the generation of multiple recombinant viruses at a time was established.