• Journal of the Korean Wood Science and Technology
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• v.34 no.1
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• pp.68-78
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• 2006

The purpose of this study was to estimate the identification of thermophilic hacterial with fermentation of pig manure. To identify the characters of thermophilic bacteria related to fermentation at a high temperature condition, we selected 28 different kinds of original settling thermophilic bacteria that were sampled at different 23 areas. They were distributed 1$1{\times}10^5{\sim}10^8CFU$ at medium and the enzyme activity at $55^{\circ}C$ incubation condition, especially cellulase and a-amylase, were higher than those of $30^{\circ}C$. Partial sequencing data for 165 rDNA region were obtained from 28 samples representing 15 different genera. Bacillus subcilis, one of those bacteria, has endodermic spores at high fermented condition.

• KSBB Journal
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• v.28 no.4
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• pp.217-224
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• 2013

A bacterium CGH18 exhibiting antioxidizing and chitin-degrading activities in the colloidal chitin culture medium was isolated from salt-fermented crab. This strain was identified as Bacillus idriensis based on 16S rDNA sequence homology search. Its crude extract was partitioned between n-BuOH and $H_2O$. The organic layer was further partitioned between $CH_2$ $Cl_2$ and $H_2O$. Antioxidant activities of crude extract and its solvent fractions were evaluated using five different bioassay methods, including the degree of occurrence of intracellular reactive oxygen species (ROS), peroxynitrite scavenging (ONOO), and oxidative damage of genomic DNA. All fractions exhibited significant antioxidant activity in bioassay systems used.

• Journal of the korean academy of Pediatric Dentistry
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• v.25 no.2
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• pp.292-303
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• 1998

The arbitrary primer polymerase chain reaction(AP-PCR) and Southern blot restriction fragment length polymorphism(RFLP) were used to genotype the cariogenic pathogen S. mutans in children. Following the morphologic chracteristics of colony on selective medium for S. mutans, total genomic DNA from 155 strains was extracted by conventional methods. Among 155 strains, 143 strains (92.3%) were confirmed S. mutans by PCR with dexA gene and 114 strains were used in this study. Three random sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 2% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI and BamHI, separated on a 0.7 % agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterised 1.6 kilobases (kb) DNA fragment cloned from gtf B gene of S. mutans. The probe was labeled with isotope[$^{32}P-{\alpha}CTP$], and hybridized fragments were detected with intensifying screen. AP-PCR produced 4-8 DNA bands in the 0.25-10 kb regions and distinguished 9, 10 or 12 genotypes, depending on the specific primer used. Southern blot RFLP analysis revealed 2 hybridization patterns consisting of 1 DNA fragments 450, 500 bp. These results indicate that AP-PCR is more discriminative method for genotyping of S. mutans.

• Korean journal of applied entomology
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• v.38 no.2
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• pp.145-149
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• 1999

To develop the baculovirus expression vector system (BEVS) adopting p10 gene promoter of Spodoptera exigua nucleopolyhedrovirus (SeNPV), we characterized the p10 gene of SeNPV. The nucleotide sequence of 545 bases including the coding region of p10 gene was determined. Compared with the previously reported SeNPV p10 gene (Zuidema et al., 1993), 4 bases were different in the 5' and 3' flanking region but no difference was found in the coding region. The p10 gene was located within Xho I 1.5 Kb, Sph 1 2.4 Kb and Cla I 4.0 Kb fragments by Southern hybridization analysis. Also, the Sph I 2.4 Kb and the Cla I 4.0 Kb fragments were cloned and their restriction enzyme maps were determined.

• Animal cells and systems
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• v.1 no.1
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• pp.129-134
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• 1997

HLA-A2 is one of the most diversified HLA-class I antigen with 17 subtypes so far identified at the molecular level. HLA-A*02 subtyping has significant implications on the tissue typing for organ and bone marrow transplantations. Recently, DNA-based typing methods have been successfully applied to the elucidation of HLA gene polymorphisms. In the present study, HLA-A*O2 genotyping was established by using nested polymerase chain reaction-sequence specific primers (PCR-SSP) and distribution of A*O2 alleles were determined in Korean individuals. Genomic DNA prepared from four B-lymphoblastoid cell lines and lymphocytes from serologically defined 48 HLA-A2 Korean individuals by phenol/chloroform extractions was typed. The results of the four B-lymphoblastoid cells were consistent with the previous data typed by PCR analysis. Five A*O2 alleles-A*0201, A*0203, A*0206, A*0207 and A*0210-were commonly observed in a total of 17 A*02 alleles. Of these, A*0207 (f=49.0%) was the most frequent allele in Korean population. A*0206 (f=28.3%) and A*0201 (f=17.0%) were also found frequently while A*0203 and A*0210 types were observed in less than 5%. In conclusion, the high level of discrimination for HLA-A*O2 alleles will prove useful and informative in the study of transplant survival, and may identify the importance of allelic differences not readily detectable by serology on host and donor compatibility.

• Animal cells and systems
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• v.5 no.3
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• pp.163-177
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• 2001

Most, if not all, aphids harbor intracellular bacterial symbionts, called Buchnera, in their bacteriocytes, huge cells differentiated for this purpose. The association between Buchnera and aphids is so intimate, mutualistic and obligate that neither of them can any longer reproduce independently. Buchnera are vertically transmitted through generations of the host insects. Evidence suggests that Buchnera were acquired by a common ancestor of aphids 160-280 million years ago, and have been diversified, since then, in parallel with their aphid hosts. Molecular phylogenetic analyses indicate that Buchnera belong to the g subdivision of the Proteobacteria. Although Buchnera are close relatives of Escherichia coli, they contain move than 100 genomic copies per cell, and their genome size is only one seventh that of E. coli. The complete genome sequence of Buchnera revealed that their gene repertoire is quite different from those of parasitic bacteria such as Mycoplasma, Rickettsia and Chlamydia, though their genome sizes have been reduced to a similar extent. Whereas these parasitic bacteria have lost most genes for the biosynthesis of amino acids, Buchnera retain many of them. In particular, Buchnera's gene repertoire is characteristic in the richness of the genes for the biosynthesis of essential amino acids that the eukaryotic hosts are not able to synthesize, reflecting a nutritional role played by these symbionts. Buchnera, when housed in the bacteriocyte, selectively synthesize a large amount of symbionin, which is a homolog of GroEL, the major stress protein of E. coli. Symbionin not only functions as molecular chaperone, like GroEL, but also has evolutionarily acquired the phosphotransferase activity through amino acid substitutions. Aphids usually profit from Buchnera's fuction as a nutritional supplier and, when faced with an emergency, consume the biomass of Buchnera cells as nutrient reserves.

• The Korean Journal of Mycology
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• v.39 no.3
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• pp.180-184
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• 2011

This study was carried out to identify the genetic characteristics among isolates of Paecilomyces spp.and Cordyceps spp. by URP-PCR analysis. Twenty URP (universal rice primer) primers of 20 mer which were designed from repetitive sequence of rice, were used for producing PCR DNA fingerprints of the mushrooms. Of them, 5 URP primers, URP2F, URP2R, URP9F, URP4R, and URP17R amplified genomic DNA of the mushrooms with polymorphic PCR patterns. On isolates of Cordyceps militaris, primers URP1F, URP2R, URP6R and URP17R produced PCR polymorphic bands of 4 types. Isolates of Cordypces sp. that are isolated from different area of Korea were identical to isolate of C. militaris, while other species of Cordypces were different to the PCR profiles. However, the URP primers did not identify the polymorphism of PCR profile on isolates of P. japonica.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.49-49
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• 2014

In order to find indigenous beneficial fungal species from crop field soils of Nigeria, 23 soil samples were collected from various places of Nigeria in June, 2013 and fungi were isolated through serial dilution technique. Isolated fungi were purified and differentiated according to their morphological and microscopic characteristics. In total, 38 different representative isolates were recovered and the genomic DNA of each isolates was extracted using QIAGEN$^{(R)}$ Plasmid Mini Kit (QIAGEN Sciences, USA) and the identification of fungi was carried out by sequence analysis of internal transcribed spacer (ITS) region of the 18S ribosomal DNA (18S rDNA). Recovered isolates belonged to 9 fungal genera comprising Fusarium, Aspergillus, Chaetomium, Coniothyrium, Dipodascaceae, Myrothecium, Neosartorya, Penicillium and Trichoderma. Aspergillus spp., Penicillium spp. and Trichoderma spp. were the most dominant taxa in this study. The antagonistic potentiality of species belonged to Trichoderma against 10 phytopathogenic fungi (F. oxysporum, C. gloesporoides, P. cytrophthora, A. alternata, A. solani, S. rolfsii, F. solani, R. solani, S. sclerotiorum and P. nicotiana) was assessed in vitro using dual culture assay. The dual culture assay results showed varied degree of antagonism against the tested phytopathogens. The potential Trichoderma spp. will be further evaluated for their antagonistic and plant growth promotion potentiality under in vivo conditions.

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.459-466
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• 2013

The lactic acid bacterium Streptococcus thermophilus is widely used as a starter culture for the production of dairy products. Whole-genome sequencing is expected to utilize the genetic basis behind the metabolic functioning of lactic acid bacterium (LAB), for development of their use in biotechnological and probiotic applications. We sequenced the whole genome of Streptococcus thermophilus MTCC 5461, the strain isolated from a curd source, by 454 GS-FLX titanium and Ion Torrent PGM. We performed comparative genome analysis using the local BLAST and RDP for 16S rDNA comparison and by the RAST server for functional comparison against the published genome sequence of Streptococcus thermophilus CNRZ 1066. The whole genome size of S. thermophilus MTCC 5461 is of 1.73Mb size with a GC content of 39.3%. Streptococcal virulence-related genes are either inactivated or absent in the strain. The genome possesses coding sequences for features important for a probiotic organism such as adhesion, acid tolerance, bacteriocin production, and lactose utilization, which was found to be conserved among the strains MTCC 5461 and CNRZ 1066. Biochemical analysis revealed the utilization of 17 sugars by the bacterium, where the presence of genes encoding enzymes involved in metabolism for 16 of these 17 sugars were confirmed in the genome. This study supports the facts that the strain MTCC 5461 is nonpathogenic and harbors essential features that can be exploited for its probiotic potential.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2010.10a
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• pp.11-11
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• 2010

The Korean ginseng, Panax ginseng C. A. Meyer is a popular medicinal herb in Araliaceae. Genetic map in crops provides valuable information for breeding, genetic and genomic researches. However, little information is available for construction of genetic map in ginseng. Up to now, we have produced large amounts of expressed sequence tags (ESTs) from four ginseng cultivars (37Mb, 49Mb, 39Mb, 47Mb from Gopoong, Gumpoong, Chunpoong and Yunpoong respectively using pyrosequencing technique and 5Mb from normalized full-length cDNA library of Chunpoong) to obtain comprehensive information of gene expression, and constructed EST database including ESTs from public database. Till now, we designed 261 SSR primer sets using EST sequences and identified 106 intergenic polymorphic markers. And 44 of the 106 showed polymorphisms among panax ginseng cultivars. Among 44 markers, 27 SSR polymorphic markers were inspected to 51 $F_2$ population from Yunpoong x Chunpoong, which showed good at the fitness of Mendellian segregation ratio 1:2:1. To enrich the number of markers, and thus construct high resolution genetic map which can be used as frame map for further genome sequencing. we are planning to develop large scale EST-derived SNP markers which are available in the F2 population. This study provides genetic information as well as foundation for ginseng researches such as genetics, genomics, breeding, and the final goal for whole genome sequencing. This study was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea (Grant No. 609001-051SB210).