• Journal of Wetlands Research
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• v.22 no.2
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• pp.100-105
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• 2020

Northern pipevine (Aristolochia contorta) commonly inhabits marginal areas between waterside and terrestrial vegetation. In particular, A. contorta is ecologically important in the marginal areas as a food plant of dragon swallowtail butterfly (Sericinus montela), which is designated as vulnerable species in the Republic of Korea. For long-term sustainability of the plant population, assessment of the genetic diversity of exist populations should be conducted. Genomic DNA of A. contorta leaf samples were extracted from four populations where the vigorous growth were observed in the South Korea. Intra-population genetic diversity and inter-population genetic distance were assessed using randomly amplified polymorphic DNA (RAPD) with five polymorphic random primers. Overall genetic diversity was lower, compared to other wetland species (h: 0.0607 ~ 0.1401; I: 0.0819 ~ 0.1759), while GP showed the highest intra-population genetic diversity. Despite of the geographical distance, GP showed the larger genetic distance from other populations. This result seemed to be caused by the fragmented habitat and lower sexual reproduction of A. controta. Mixture of the different source populations and construction of the proper environmental condition such as shade and physical support for sexual reproduction should be considered for conservation of A. contorta population.

• The Korean Journal of Mycology
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• v.36 no.2
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• pp.130-137
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• 2008

To distinct the commercial strains in Pleurotus spp., 81 strains in eight Pleurotus species were used. DNA fingerprinting using URP-PCR was conducted to determine the phylogenetic relationships among Pleurotus strains. DNA profiles of Pleurotus species obtained by twelve URP primers were analyzed for genetic similarity by NTSYS program. We could divide strains into ten clusters, in which three of them belong to P. ostreatus and the others to the different species, respectively. At the 76% similarity level, 70 P. ostreatus strains were distinguished into three clusters. Cluster I contained 35 strains and some of them showed almost 100% similarity, one strain closely related to Weonhyeong and six strains closely related to Wangheukpyeong. In cluster II, twenty-one out of 23 strains showed 100% to Suhan. Cluster III contained twelve strains, including six strains closely related to Chunchu-2. The results suggested that there are many same strains with different names in mushroom spawn market.

• Korean Journal of Microbiology
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• v.44 no.1
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• pp.74-79
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• 2008

The purpose of this study is to clone cgt gene from Paenibacillus sp. JB-13 and to overexpress the protein in E. coli. For this purpose, the cgt gene was amplified from Paenibacillus sp. JB-13 genomic DNA by PCR using degenerate oligonucleotide primers. The sequence analysis results showed that the cgt gene from Paenibacillus sp. JB-13 has 98% homology with the cgt gene of Bacillus sp. To overexpress the protein, the cgt gene was cloned into pEXP7 expression vector and transformed into E. coli. The production of CGTase by recombinant E. coli was optimized under following conditions: 0.5% glucose, 3.0% polypeptone, 0.3% $K_2HPO_4$, 0.5% NaCl, and 7.0 of initial pH, 2.0% of inoculum, $37^{\circ}C$ of culture temperature for 14 hr. And the optimal agitation was found at 0.1 vvm. The synthesis of 2-O-${\alpha}$-D-Glucopyranosyl L-Ascorbic acid (AA-2G) using the CGTase expressed in E. coli was identified as AA-2G by HPLC and HPLC confirmed that treating AA-2G made by cloned CGTase with ${\alpha}$-glucosidase substantially produced AA and glucose.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.81-89
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• 2004

Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and lymphotoxin-$\alpha$ (LT-$\alpha$, TNF-$\beta$) can initiate and perpetuate human diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA), and insulin-dependent diabetes mellitus (IDDM). TNFs can be blocked by the use of soluble TNF receptors. However, since monomeric soluble receptors generally exhibit low affinity or function as agonists, the use of monomeric soluble receptors has been limited in the case of cytokines such as TNF-$\alpha$, TNF-$\alpha$, interleukin (IL)-1, IL-4, IL-6, and IL-13, which have adapted to a multi component receptor system. For these reasons, very high-affinity inhibitors were created for the purpose of a TNFs antagonist to bind the TNFR and trigger cellular signal by using the multistep polymerase chain reaction method. First, recombinant simple TNFR-Ig fusion proteins were constructed from the cDNA sequences encoding the extracellular domain of the human p55 TNFR (CD120a) and the human p75 TNFR (CD120b), which were linked to hinge and constant regions of human $IgG_1$ heavy chain, respectively using complementary primers (CP) encoding the complementary sequences. Then, concatameric TNFR-Ig fusion proteins were constructed using recombinant PCR and a complementary primer base of recombinant simple TNFR-Ig fusion proteins. For high level expression of recombinant fusion proteins, Chinese hamster ovary (CHO) cells were used with a retroviral expression system. The transfected cells produced the simple concatameric TNFR-Ig fusion proteins capable of binding TNF and inactivating it. These soluble versions of simple concantameric TNFR-Ig fusion proteins gave rise to multiple forms such as simple dimers and concatameric homodimers. Simple TNFR-1g fusion proteins were shown to have much more reduced TNF inhibitory activity than concatameric TNFR-Ig fusion proteins. Concatameric TNFR-Ig fusion proteins showed higher affinity than simple TNFR-Ig fusion proteins in a receptor inhibitor binding assay (RIBA). Additionally, concatameric TNFR-Ig fusion proteins were shown to have a progressive effect as a TNF inhibitor compared to the simple TNFR-Ig fusion proteins and conventional TNFR-Fc in cytotoxicity assays, and showed the same results for collagen induced arthritis (CIA) in mice in vivo.

• Journal of Plant Biotechnology
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• v.29 no.4
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• pp.229-233
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• 2002

Ferritin is ubiquitous in bacteria, animals and plants. Ferritin is thought to play two main roles in living cells to provide iron for the synthesis of iron protein such as ferretoxin and cytochromes and to prevent damage from radicals produced by iron/dioxygen interaction. To enhance the resistance of potato to Erwinia carotovora, the soybean ferritin gene was introduced into the potato either under CaMV 35S or hsr203J promoter. Potato transgenic plants were screened by PCR analysis using specific primers to the ferritin gene. Expression of ferritin gene under CaMV 35S and hsr203J promoter in potato transgenic plants was confirmed by northern blot analysis. hsr203J promoter known to pathogen inducible in tobacco drives the induction upon Phytophthora infestan in potato and the transcript level of ferritin gene was extremely high after 24 hours post inoculation. One of transformants under CaMV 35S promoter was increased 2.5 fold than untransformant. Each one of transgenic potato containing gene promoter CaMV 35S and hsr203J-ferrtin fusion exhibited tolerance against potato soft rot.

• Journal of Food Hygiene and Safety
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• v.28 no.2
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• pp.181-187
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• 2013

Identification of main ingredients in starches has been investigated using physicochemical analysis method mainly. However, physicochemical properties such as particle size have limitations in determining the differences among mixed starches. Therefore, we developed a molecular biological method to identify materials used in starch, as a sample, 11 kinds of starches including sweet potato starch, potato starch, corn starch, and tapioca starch. DNeasy plant mini kit, magnetic DNA purification system, and CTAB methods were used to extract DNA from samples. After gene extraction, whole genome amplification (WGA) was performed to amplify the extracted DNA. Species-specific primers were used as followings: ib-286-F/ib-286-R (105 bp), Pss 01n-5'/Pss 01n-3' (216 bp), SS11b 3-5'/SS11b 3-3' (114 bp), and SSRY26-F/SSRY26-R (121 bp) gene for sweet potato, potato, corn, and tapioca, respectively. In this study, we could confirm the main ingredients using WGA and PCR method.

• The Korean Journal of Mycology
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• v.40 no.1
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• pp.7-10
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• 2012

Endophytic fungi were isolated from the roots of halophytes, Suaeda japonica and Carex scabrifolia in the Suncheonbay. The ITS region in rDNA of 15 endophytic fungal strains were amplified using PCR with universal primers ITS1 and ITS4, and those amplified fragments were sequenced. Based on ITS sequence, five fungal genera were identified in S. japonica and seven fungal genera were identified in C. scabrifolia. The Shannon's diversity index (H') of endophytic fungi isolated from S. japonica and C. scabrifolia was 1.561 and 1.889, respectively. In phylogenetic analysis, it was shown that Ascomycota and Pezizomycotina was widely distributed both in S. japonica and C. scabrifolia. Also, Sordariomycetes, Dothideomycetes and Eurotiomycetes were shown to be distributed in these halophytes used in this experiment.

• Research in Plant Disease
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• v.21 no.3
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• pp.235-242
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• 2015

A 2014 nationwide survey in radish fields investigated the distribution of common viruses and possible emerging viruses. Radish leaves with virus-like symptoms were collected and 108 samples assayed by RT-PCR using specific primers for Radish mosaic virus (RaMV), Cucumber mosaic virus (CMV), and Turnip mosaic virus (TuMV); 47 samples were TuMV positive, and RaMV and CMV were detected in 3 and 2 samples, respectively. No samples showed double infection of TuMV/RaMV, or RaMV/CMV, but two double infections of TuMV/CMV were detected. TuMV isolates were sorted by symptom severity, and three isolates (R007-mild; R041 and R065-severe) selected for BLAST and phylogenetic analysis, which indicated that the coat protein (CP) of these isolates (R007, R041, and R065) have approx. 98-99% homology to a previously reported TuMV isolate. RaMV CP showed approx. 99% homology to a previously reported isolate, and the CMV CP is identical to a previously reported Korean isolate (GenBank : GU327368). Three isolates of TuMV showing different pathogenicity (degree of symptom severity) will be valuable to study determinants of pathogenicity.

• Journal of Plant Biotechnology
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• v.39 no.3
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• pp.196-204
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• 2012

This study was carried out to develop reliable systems for somatic embryogenesis in oil palm tree (Elaeis guineensis Jacq.), and to verify the somaclonal variants by RAPD analysis. Embryogenic callus was induced successfully on modified half-strength MS medium containing $NaH_2PO_4{\cdot}2H_2O$ and casein. Embryogenic callus was further developed to somatic embryo mass (SEM), which is very hard and bonded tightly each other. Plantlets were proliferated when SEM was cultured on modified MS medium containing half strength $NH_4NO_3$, casein and L-ascorbic acid. Plantlets were transplanted into pots containing artificial soils. When RAPD analysis was conducted using randomly selected 95 in vitro plantlets and 19 random primers, somaclonal variation was detected using BNR35 primer. There was missing band around 1 kb in #22, #28, #35, and #77 plantlets. In addition, bands obtained from #28, #35, and #77 was much stronger than other normal bands. The blast results at NCBI revealed that somaclonal variation observed in this study was related to chloroplast genome of oil palm. The results also revealed that oil palm reproduction system through somatic embryogenesis is quite reliable and early detection of somaclonal variants seem to be possible at in vitro stage by RAPD analysis.

• Korean Journal of Medicinal Crop Science
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• v.11 no.2
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• pp.102-108
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• 2003

This experiment was conducted to evaluate the diversity and purity of the Korean ginseng (Panax gjnseng) lines developed by the pure line selection using RAPD markers. Four primer (OPA 19, OPM 11, URP 3 and UBC 98) out of the 48 primer tested produced band which showed within-line polymorphisms at least in one line. Within-line polymorphisms were detected in six lines by OPA 19, in four lines by URP 03, in five lines by OPM 11, and in one line by UBC 98 respectively. Five plants obtained from the commercial cultivar 'Cheonpung' were differentiated using the primers OPA 19 and OPM 11. Five plants obtained from the 'Yeonpung were differentiated using the primer OPM 11. Detection of within-line RAPD polymorphisms might be attributed to the fact that cross pollination appear in P. gjnseng and a long period of three to four years required to reach the reproductive stage thereby delay the process to homozygosity.