• Journal of Life Science
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• v.33 no.10
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• pp.808-819
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• 2023

This study was conducted to develop a selection marker for the identification of the Bacillus licheniformis K12 strain in microbial communities. The strain not only demonstrates good growth at moderate temperatures but also contains enzymes that catalyze the decomposition of various polymer materials, such as proteases, amylases, cellulases, lipases, and xylanases. To identify molecular markers appropriate for use in a microbial community, a search was conducted to identify variable gene regions that show considerable genetic mutations, such as recombinase, integration, and transposase sites, as well as phase-related genes. As a result, five areas were identified that have potential as selection markers. The candidate markers were two recombinase sites (BLK1 and BLK2), two integration sites (BLK3 and BLK4), and one phase-related site (BLK5). A PCR analysis performed with different Bacillus species (e.g., B. licheniformis, Bacillus velezensis, Bacillus subtilis, and Bacillus cereus) confirmed that PCR products appeared at specific locations in B. licheniformis: BLK1 in recombinase, BLK2 in recombinase family protein, and BLK3 and BLK4 as site-specific integrations. In addition, BLK1 and BLK3 were identified as good candidate markers via a PCR analysis performed on subspecies of standard B. licheniformis strains. Therefore, the findings suggest that BLK1 can be used as a selection marker for B. licheniformis species and subspecies in the microbiome.

• Korean Journal of Microbiology
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• v.43 no.4
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• pp.256-263
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• 2007

We performed the biochemical characteristics, molecular epidemiologocal analysis, and drug susceptibility test on V. vulnificus isolated from environmental sources in Incheon. For this study, 233 strains were isolated from seawater, sediment, shellfish. V. vulnificus isolates were divided into 15 biochemical groups, which were positive for ONPG and Amygdalin test. Among the 209 strains, 206 (98.6%) strains and 110 (52.6%) strains revealed positive for vvhA and viuB gene, and the viuB gene detection rates of V. vulnificus from seawater, shellfish and sediment were 48%, 48.5% and 61.6%, respectively. From disc diffusion test on 175 isolates, most of strains were sensitive to Imipenem (100.0%), Sulfamethoxazole/trimethoprim (98.9%), Tetracycline, Ciprofloxacin (98.3%), Ampicillin/sulbactam (97.1%), Ohloramphenicol (96.6%), Cefepime (94.9%) and Ceftriaxone (94.8%), multi-drug resistance rates was 31.5% of seawater, 34.4% of sediment and 29.2% of shellfish. PFGE was performed on 233 V. vulnificus isolates with the objective of investigating the extent of genetic diversity of these isolates in our region. We could find that at least 126 different PFGE patterns were generated according by 90% of similarity and 13 clusters by 58% of similarity. The major cluster was type I (44.6%) during the most of the year, and type J was frequent pattern in June and October. There were 9 distinct PFGE types in July, 8 types in August, 7 types in June, 6 types in September, 5 types in October 3 types in May and 1 type in March.

• Restorative Dentistry and Endodontics
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• v.30 no.5
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• pp.409-422
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• 2005

The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 XPBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing $5\%$ sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions, This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.183-188
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• 2015

This study analyzed the coat color-related genes of MC1R, ASIP, ECA3-inversion, and STX17 of 1,462 Jeju horses administered by the Jeju Special Self-Governing Province. This was done to investigate the distributional characteristics of coat color-related genes in the Jeju horse group and the changes of its coat color-related genes by generation. The genotype frequency of the MC1R gene of $E^+/E^+$ and $E^+/E^e$ related to black coat color was 0.122 and 0.447, respectively, while $E^e/E^e$ of the chestnut genotype was 0.429. The genotype frequency of the ASIP gene of $A^A/A^A$, $A^A/A^a$, and $A^a/A^a$ was 0.46, 0.448, and 0.091, respectively, where the genotype frequency of $A^a/A^a$ turned out to be relatively low. The To/To and +/To genotype that manifests the Tobiano shape was 0.001 and 0.119, respectively, with the share of Tobiano shape around 12%. The genotype frequency of G/G and G/g of STX17 related to grey coat color was 0.002 and 0.680, respectively, with the share of grey horses among the Jeju horse group at 68.2%. As for the change of coat color genes by generation, no large changes were observed in the MC1R and ASIP genes. In ECA3-inversion, the To allele that manifests Tobiano significantly decreased following the generational change (p<0.05), while the STX17 G allele related to grey coat color significantly increased following the generational change (p<0.05). It will be necessary to examine the coat color genes when selecting breeding horses so that the diversity of coat colors among the Jeju horse group can be maintained.

• Korean Journal of Plant Resources
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• v.32 no.5
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• pp.519-542
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• 2019

Twenty soybean cultivars developed recently were assessed using 27 insertion and deletion (InDel) markers derived from dense variation blocks (dVBs) of soybean genome. The objective of this study is to identify the distinctness and genetic relationships among a total of 169 soybean accessions including new cultivars. The genetic homology between 149 accessions in the soybean barcode system and 20 new cultivars was 61.3% on average with the range from 25.9% to 96.3%, demonstrating the versatile application of these markers for cultivars identification. The phylogenic analysis revealed four subgroups related to their usage. The 80% of cultivars for vegetable and early maturity and the 65.9% of cultivars for bean sprouts were clustered in subgroup I-2 and II-2, respectively, indicating of the limited gene pools of their crossing parents in breeding. On the other hands, the cultivars for soy sauce and tofu with considerable gene flow by genome reshuffling were distributed evenly to several subgroups, I-1 (44.4%), I-2 (26.4%) and II-2 (23.6%). We believe that the 27 InDel markers specific to dVBs can be used not only for cultivar identification and genetic diversity, but also in breeding purposes such as introduction of genetic resources and selection of breeding lines with target traits.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.65 no.3
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• pp.199-213
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• 2020

Grain size and shape are the two important components contributing to rice yield and quality. To analyze traits related to grain-shape, a total of 272 varieties derived from japonica, japonica black and Tongil-type rice accession in Korea were evaluated in this study. The traits, grain length (GL), grain width (GW), grain thickness (GT), length to width ratio (RLW), and 1000-grain weight (TGW) were measured and replicated 10 times. Genes (GW2, GS3, qGL3, qSW5, GS5, TGW6, GW7, and GW8) related to grain-shape were validated in the accessions using specific DNA marker sets. K-mean clustering of the accession based on phenotypic data revealed three groups: group 1 was classified by GW and GT and included most of japonica type, group 2 was classified by RLW and GL reached a medium size and possessed a half spindle-shaped type, and group 3 was classified by TGW, reached a long size and possessed a semi-round shape. In validation tests using the marker sets, both gw2 and tgw6 were validated in less than 1% of the tested accessions and two allelic types, qgl3 and gw8, were only verified in Tongil-type accessions. For GW8 and GW2, any different amplicons were not amplified in any japonica or Tongil-type accessions, respectively. In order to suggest the representative grain-shape gene combinations for each ecotype, the allelic combinations were evaluated by PCR analysis. Cj1 and 2 in japonica (Cj1-7), Cj_b1 and 2 in japonica-black (Cj_b1-3), and CT3 in Tongil-type (CT1-13) turned out to be the dominant combination in each ecotype, respectively. In addition, the results revealed that introgression of four genes (gw2, gs3, qSW5, and GS5) would expand the diversity of grain shape in Korean japonica varieties. The gene combinations information could be utilized practically to understand or enhance grain shape in japonica rice breeding program.

• Natural Product Sciences
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• v.8 no.1
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• pp.1-15
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• 2002

An International Cooperative Biodiversity Group (ICBG) program based at the University of Illinois at Chicago initiated its activities in 1998, with the following specific objectives: (a) inventory and conservation of of plants of Cuc Phuong National Park in Vietnam and of medicinal plants of Laos; (b) drug discovery (and development) based on plants of Vietnam and Laos; and (c) economic development of communities participating in the ICBG project both in Vietnam and Laos. Member-institutions and an industrial partner of this ICBG are bound by a Memorandum of Agreement that recognizes property and intellectual property rights, prior informed consent for access to genetic resources and to indigenous knowledge, the sharing of benefits that may arise from the drug discovery effort, and the provision of short-term and long-term benefits to host country institutions and communities. The drug discovery effort is targeted to the search for agents for therapies against malaria (antimalarial assay of plant extracts, using Plasmodium falciparum clones), AIDS (anti-HIV-l activity using HOG.R5 reporter cell line (through transactivation of the green fluorescent protein/GFP gene), cancer (screening of plant extracts in 6 human tumor cell lines - KB, Col-2, LU-l, LNCaP, HUVEC, hTert-RPEl), tuberculosis (screening of extracts in the microplate Alamar Blue assay against Mycobacterium tuberculosis $H_{37}Ra\;and\;H_{37}Rv),$ all performed at UIC, and CNS-related diseases (with special focus on Alzheimer's disease, pain and rheumatoid arthritis, and asthma), peformed at Glaxo Smith Kline (UK). Source plants were selected based on two approaches: biodiversity-based (plants of Cuc Phuong National Park) and ethnobotany-based (medicinal plants of Cuc Phuong National Park in Vietnam and medicinal plants of Laos). At mc, as of July, 2001, active leads had been identified in the anti-HIV, anticancer, antimalarial, and anti- TB assay, after the screening of more than 800 extracts. At least 25 biologically active compounds have been isolated, 13 of which are new with anti-HIV activity, and 3 also new with antimalarial activity. At GSK of 21 plant samples with a history of use to treat CNS-related diseases tested to date, a number showed activity against one or more of the CNS assay targets used, but no new compounds have been isolated. The results of the drug discovery effort to date indicate that tropical plant diversity of Vietnam and Laos unquestionably harbors biologically active chemical entities, which, through further research, may eventually yield candidates for drug development. Although the substantial monetary benefit of the drug discovery process (royalties) is a long way off, the UIC ICBG program provides direct and real-term benefits to host country institutions and communities.

• Journal of Animal Science and Technology
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• v.46 no.6
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• pp.917-924
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• 2004

Using PCR-RFLP haplotyping for the mitochondrial DNA(mtDNA) fragment containing the NADH dehydrogenase 2 gene(ND2) and three tRNA genes(tRNA-Met, tRNA-Trp and tRNA-Ala), we characterized the genetic diversity of five pig breeds including Jeju native pigs. mtDNA polymorphisms showing distinct cleavage patterns were found in the pig breeds. Two digestion patterns were detected when HaeIII- and Hinfl-RFLP, and four in the Tsp5091-RFLP analyses. Combining the three restriction enzyme digestion patterns found in five different pig breeds, four mtDNA haplotypes were observed and the haplotype frequencies were significantly different by the pig breeds. A monomorphic haplotype, mtWB, was observed in both Korean wild boars and Large White pigs. Both Duroc and Landrace pigs contained two haplotypes suggesting their multiple maternal lineages. Jeju native pig has two haplotypes(mtJN and mtJD). Of these, mtJN is identified as a Jeju native pig specific haplotype. This study suggested that more than two progenitor populations have been taken part in the domestication process of the Jeju native pig population, and/or probably subsequent crossing with other pig breeds from near east Asia. Unlike with our prediction, there was no direct evidence under molecular levels on the maternal introgression of Korean wild boar in the domestication of Jeju native pigs. In conclusion, specificity of mtDNA haplotypes related to pig breeds win be useful for identifying the maternal lineage as wen as constructing the genealogical pedigree in pigs.

• Journal of Dairy Science and Biotechnology
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• v.34 no.4
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• pp.203-216
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• 2016

Probiotic microorganisms are thought to provide health benefits when consumed. In 2001, the World Health Organization defined probiotics as "live microorganisms which confer a health benefit on the host, when administered in adequate amounts." Three methods for screening potential probiotics have currently widely available. (1) In vitro assays of potential probiotics are preferred because of their simplicity and low cost. (2) The use of in vivo approaches for exploring various potential probiotics reflects the enormous diversity in biological models with various complex mechanisms. (3) Potential probiotics have been analyzed using several genetic and omics technologies to identify gene expression or protein production patterns under various conditions. However, there is no ideal procedure for selecting potential probiotics than testing cadidate strains on the target population. Hence, in this review, we provide an overview of the different methodologies used to identify new probiotic strains. Furthermore, we describe futre perspectives for the use of in vitro, in vivo and omics in probiotic research.

• Proceedings of the Ginseng society Conference
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• 1978.09a
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• pp.149-157
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• 1978

Ginseng is the English common name for the species in the genus Panax. This article gives a broad botanical review including the morphological characteristics, ecological amplitude, and the ethnobotanical aspect of the genus Panax. The species of Panax are adapted for life in rich loose soil of partially shaded forest floor with the deciduous trees such as linden, oak, maple, ash, alder, birch, beech, hickory, etc. forming the canopy. Like their associated trees, all ginsengs are deciduous. They require annual climatic changes, plenty of water in summer, and a period of dormancy in winter. The plant body of ginseng consists of an underground rhizome and an aerial shoot. The rhizome has a terminal bud, prominent leafscars and a fleshy root in some species. It is perennial. The aerial shoot is herbaceous and annual. It consists of a single slender stem with a whorl of digitately compound leaves and a terminal umbel bearing fleshy red fruits after flowering. The yearly cycle of death and renascence of the aerial shoot is a natural phenomenon in ginseng. The species of Panax occur in eastern North America and eastern Asia, including the eastern portion of the Himalayan region. Such a bicentric generic distributional pattern indicates a close floristic relationship of the eastern sides of two great continental masses in the northern hemisphere. It is well documented that genera with this type of disjunct distribution are of great antiquity. Many of them have fossil remains in Tertiary deposits. In this respect, the species of Panax may be regarded as living fossils. The distribution of the species, and the center of morphological diversification are explained with maps and other illustrations. Chemical constituents confirm the conclusion derived from morphological characters that eastern Asia is the center of species concentration of Panax. In eastern North America two species occur between longitude $70^{\circ}-97^{\circ}$ Wand latitude $34^{\circ}-47^{\circ}$ N. In eastern Asia the range of the genus extends from longitude $85^{\circ}$ E in Nepal to $140^{\circ}$ E in Japan, and from latitude $22^{\circ}$ N in the hills of Tonkin of North Vietnam to $48^{\circ}$ N in eastern Siberia. The species in eastern North America all have fleshy roots, and many of the species in eastern Asia have creeping stolons with enlarged nodes or stout horizontal rhizomes as storage organs in place of fleshy roots. People living in close harmony with nature in the homeland of various species of Panax have used the stout rhizomes or the fleshy roots of different wild forms of ginseng for medicine since time immemorial. Those who live in the center morphological diversity are specific both in the application of names for the identification of species in their communication and in the use of different roots as remedies to relieve pain, to cure diseases, or to correct physiological disorders. Now, natural resources of wild plants with medicinal virtue are extremely limited. In order to meet the market demand, three species have been intensively cultivated in limited areas. These species are American ginseng (P. quinquefolius) in northeastern United States, ginseng (P. ginseng) in northeastern Asia, particularly in Korea, and Sanchi (P. wangianus) in southwestern China, especially in Yunnan. At present hybridization and selection for better quality, higher yield, and more effective chemical contents have not received due attention in ginseng culture. Proper steps in this direction should be taken immediately, so that our generation may create a richer legacy to hand down to the future. Meanwhile, all wild plants of all species in all lands should be declared as endangered taxa, and they should be protected from further uprooting so that a. fuller gene pool may be conserved for the. genus Panax.