• Title/Summary/Keyword: Chromosomal technology

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Pathogenic and Molecular Characteristics of Agrobacterium vitis strains isolated from Grapevine in Korea

  • Kim, J.G.;Kim, S.H.;Choi, J.E.;Lee, Y.K.;Kang, H.W.
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.120.2-120
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    • 2003
  • Agrobacterium vitis is a causal agent of crown-gall disease on grapevine. In Korea, grapevine variety (GeoBong) have severely been infected by the bacteria since stems of the variety were buried in soil for overwintering. Infection ratio over 70-80% was observed on 7 years old GeoBong grapevine in Ansung and Cheonan. PCR specific primers for A. vitis strains were designed using nucleotide sequences of vir A gene in Ti-Plasmid, pheA gene in chromosomal DNA and a URP-PCR polymorphic band. Three hundred bacterial strains were isolated from the different 80 galls formed on GeoBong grapevine in Cheonan and Ansung of Korea and were screened to identify A. vitis using the three specific PCR primers for Agrobacterium vitis. Twenty-four bacterial strains that are detected by the primers were further confirmed by pathogenicity and biochemical methods. To investigate the genomic diversity of the bacterial strains, twenty primers of 20 mer referred to universal rice primers (URP) were applied for PCR fingerprinting, Of them, URP2R and URP2F primers could effectively be used to detect polymorphism within the bacterial strains.

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Construction of Shuttle Promoter-probe and Expression Vectors for Escherichia coli and Bacillus subtilis, and Expression of B. thuringiensis subsp. kurstaki HD-73 Crystal Protein Gene in the Two Species

  • Park, Seung-Hwan;Koo, Bon-Tag;Shin, Byung-Sik;Kim, Jeong-Il
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.37-44
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    • 1991
  • A shuttle promoter-probe vector, pEB203, was derived from pBR322, pPL703 and pUB110. Using the vector, a useful DNA fragment, 319 bp EcoRI fragment, having strong promoter activity has been cloned from Bacillus subtills chromosomal DNA. Selection was based on chloramphenicol resistance which is dependent upon the introduction of DNA fragments allowing expression of a chloramphenicol acetyl transferase gene. The nucleotide sequence of the 319 bp fragment has been determined and the putative -35 and -10 region, ribosome binding site, and ATG initiation codon were observed. This promoter was named EB promoter and the resultant plasmid which can be used as an expression vector was named pEBP313. The crystal protein gene from B. thuringiensis subsp. kurstaki HD-73 was cloned downstream from the EB promoter without its own promoter. When the resultant plasmid, pBT313, was introduced into Escherichia coli and B. subtilis, efficient synthesis of crystal protein was observed in both cells, and the cp gene expression in B. subtilis begins early in the vegetative phase. The cell extracts from both clones were toxic to Hyphantria cunea larvae.

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Cloning and Expression of Serratia marcescens Protease Gene in Escherichia coli

  • KIM, MYUNG-HEE;SOO-KEUN CHOI;BON-TAG KOO;BYUNG-SIK SHIN;CHEON-BAE SOHN;JEONG-IL KIM
    • Journal of Microbiology and Biotechnology
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    • v.2 no.4
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    • pp.231-236
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    • 1992
  • A 5.8 kb chromosomal DNA fragment of Serratia marcescens ATCC 27117 including an extracellular serine protease gene was cloned in Escherichia coli. The cloned gene(pSMP18) caused specific excretion of the protease into the extracellular medium through the outer membrane of E. coli host cells. The protease purified from E. coli harboring pSMP18 was inactivated not by 100 mM EDTA but by 10 mM phenyl methyl sulfonyl flouride (PMSF). The molecular weight of the purified serine protease was about 66, 000 in the SDS-PAGE and the isoelectric point was approximately 5.7 in IEFㆍGel electrophoresis. The optimal pH and temperature for reaction of the purified serine protease were 9.5 and $45^\circ{C}$, respectively.

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Isolation and Functional Analysis of spy1 Responsible for Pristinamycin Yield in Streptomyces pristinaespiralis

  • Jin, Qingchao;Yin, Huali;Hong, Xiaowei;Jin, Zhihua
    • Journal of Microbiology and Biotechnology
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    • v.22 no.6
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    • pp.793-799
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    • 2012
  • A gene related to high pristinamycin yield in Streptomyces pristinaespiralis was selected by amplified fragment length polymorphism (AFLP) and its functions were investigated by gene disruption. First, a 561 bp polymorphic sequence was acquired by AFLP from high-yield recombinants compared with the S. pristinaespiralis ancestor ATCC25486, indicating that this approach is an effective means of screening for valuable genes responsible for antibiotic yield. Then, a 2,127 bp open reading frame of a gene designated spy1 that overlaps with the above fragment was identified and its structure and biological functions were investigated. In silico analysis of spy1 encoding a deduced 708-amino-acid-long serine/threonine protein kinase showed that it only contains a catalytic domain in the N-terminal region, which is different from some known homologs. Gene inactivation of chromosomal spy1 indicated that it plays a pleiotropic regulatory function in pristinamycin production, with a positive correlation to pristinamycin I biosynthesis and a negative correlation to pristinamycin II biosynthesis.

Synergistic Antifungal Activity of Phellodendri Cortex and Magnoliae Cortex against Candida albicans

  • NA, Hyunjeong;KIM, Tae-Jong
    • Journal of the Korean Wood Science and Technology
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    • v.50 no.1
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    • pp.12-30
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    • 2022
  • Many studies on plant extracts have been reported for the treatment of candidiasis caused by Candida albicans, a representative fungal infection. This study demonstrates the synergistic antifungal activity of the combination of Phellodendri Cortex and Magnoliae Cortex, previously reported to have antifungal efficacy. Considering the antifungal efficacy and the separation of the active constituents, berberine and magnolol, hot water extraction and carbon dioxide supercritical extraction were selected for Phellodendri Cortex and Magnoliae Cortex, respectively. A combination of 0.55 g/L hot water extract of Phellodendri Cortex and 0.59 g/L carbon dioxide supercritical extract of Magnoliae Cortex showed synergistic antifungal activity. The synergistic antifungal activity of 160 μM berberine and 100 μM magnolol, which are representative antifungal compounds of Phellodendri Cortex and Magnoliae Cortex, respectively, contributes to the synergistic antifungal effect of their extracts. The additive decrease in cellular ergosterol level and the increased antifungal efficacy by extracellular ergosterol suggest that disruption of the biological function of ergosterol in the cell membrane is not responsible for the synergistic antifungal activity of berberine and magnolol. Synergistic cellular release of chromosomal DNA upon mixing berberine and magnolol indicates that disruption of the cellular structure is responsible for the synergistic antifungal effect of berberine and magnolol.

Study on Genotoxicity of Crocin, a Component of Gardenia Fruit, in Bacterial and Mammalian Cell Systems

  • Choi, Hae-Yeon;Kim, Youn-Jung;Jeon, Hee-Kyung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.285-292
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    • 2008
  • Crocin is one of the major components of gardenia fruit and saffron which are widely used as natural food colorants and as traditional Chinese medicines. However, the genotoxicity data on crocin are not sufficient for safety evaluation. The purpose of this study was the examination of the genotoxicity on crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and ${H_2}{O_2}$-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay. From results, no considerable mutagenicity and clastogenicity were seen in bacteria and mammalian cells treated with crocin, by Ames test, chromosomal aberration assay, ${tk}^{+/-}$ gene forward mutation assay and comet assay. And, post-treatment with crocin significantly suppressed ${H_2}{O_2}$-induced DNA damage in a dose-dependent manner. In conclusion, the findings of the present study and other previous observations indicate that crocin has no genotoxic potential. And it showed that crocin clearly repressed the genotoxic potency of ${H_2}{O_2}$. These results suggest that anti-oxidative effects of crocin may be involved in the protective effects of DNA damage.

Identification of QTLs Affecting Physical Traits of Cooked Rice

  • Kang, Hyeon-Jung;Cho, Yong-Gu;Lee, Young-Tae;Kim, Young-Doo;Eun, Moo-Young;Shim, Jae-Uk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.1
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    • pp.70-73
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    • 1999
  • This study was conducted to ascertain the chromosomal locations and effect of quantitative trait loci (QTL) associated with the physical traits of rice (Oryza sativa L.) eating quality. One hundred sixty four recombinant inbred lines (MGRILs) of F$_{11}$ were derived from the cross between Milyang 23 (Tongil type) and Gihobyeo (japonica type). They were evaluated for six physical traits of cooked rice. Transgressive segregation was observed for all examined traits. Significant QTL were detected (LOD$\geq$2.0) in three traits, including single QTL for adhesiveness, gumminess, and chewiness of cooked rice, respectively. Phenotypic variation explained by each QTL ranged from 6.3% to 14.6%. However, no significant QTL was detected for hardness, cohesiveness, and elasticity of cooked rice. Pleiotropic effects of single QTL on different traits are observed.d.

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Colony Management and Its Application to Genetic Engineering (실험동물의 계통 보존과 유전공학에 응용)

  • 오양석
    • Journal of Embryo Transfer
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    • v.7 no.1
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    • pp.49-55
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    • 1992
  • Animal experiments need numerous kinds of animal which are suitable for every research. About 300 mouse strains are developed up to the present, but they do not give satisfaction to every researchers. So we must build up the methods of breading animals which are newly developed and of maintenance of characteristics which were developed before. To maintain experimental animal is not only proceeding the generation but also increasing the animal populations, it needs geneticai control. Genetic factors which influence to reproduction are very important to maintain colony. These factors include lethal gene, chromosomal abberation, sterility gene, etc.. With the recent development of transgenic technology, scientists now can deliberately creat numerous specific animal models. To know how to manage the colony which has genetic defect on reproduction and transgenic mice is one of the key to study in vitro fertilization.

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Discovery of Performance Traits-Linked Microsatellite Markers in Channel Catfish (Ictalurus punctatus)

  • Kim, Soon-Hag
    • Journal of Aquaculture
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    • v.18 no.2
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    • pp.130-132
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    • 2005
  • Genomics research has two ultimate applied goals: to Isolate and clone genes of economic importance for bio-technology and gene-assisted selection (GAS), and to locate and use markers for marker-assisted selection (MAS) in selective breeding programs. To this end, we have identified linked markers for feed conversion efficiency growth rate, and disease resistance to enteric septicemia of catfish (ESC). Three microsatellite markers Ip266, Ip384, and Ip607 were identified to be linked to feed conversion efficiency. Similarly one marker each was identified to be linked to growth rate (Ip607) and disease resistance to ESC (Ip477). Ip607 marker linked to both growth rate and feed conversion efficiency, indicating that the QTL for both growth rate and feed conversion efficiency may either be the same or located in the same chromosomal region in the catfish genome. On phenotypic evaluation, certain traits such as growth rate can be accurately evaluated by body weight evaluation while other traits such as disease resistance can be quite complex. The linked DNA markers will be highly useful for MAS programs and for directing further efforts of genomic mapping for important quantitative traits.

Variations in Chromosome Numbers and Saikosaponin Contents of Bupleurum falcatum L. from Different Geographical Regions

  • Son, Tae-Kwon;Kim, Kil-Ung;Jeong, Hyung-Jin;Lee, Sang-Chul
    • Plant Resources
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    • v.4 no.1
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    • pp.48-52
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    • 2001
  • Seven genetic lines of Bupleurum falcatum L. from different geographical regions were analysed for saikosaponin contents and chromosomal numbers. The somatic chromosome numbers of B. falcatum originated from Euisong, Iri, Milyang, Sangnam, Taejon, and Youngchon were 2n=20 while Mishimasaiko showed 2n=26. However, chromosome features were different in plants grown in different geographical regions. Generally, Korean lines had higher saikosaponin contents than Mishimasaiko which is Japanese and Sangnam lines had highest saikosaponin contents compared to other tested lines.

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