• Journal of Microbiology
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• v.45 no.2
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• pp.105-112
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• 2007

The objective of this study was to analyze the phylogenetic composition of bacterial community in the soil of an earth-cave (Niu Cave) using a culture-independent molecular approach. 16S rRNA genes were amplified directly from soil DNA with universally conserved and Bacteria-specific rRNA gene primers and cloned. The clone library was screened by restriction fragment length polymorphism (RFLP), and representative rRNA gene sequences were determined. A total of 115 bacterial sequence types were found in 190 analyzed clones. Phylogenetic sequence analyses revealed novel 16S rRNA gene sequence types and a high diversity of putative bacterial community. Members of these bacteria included Proteobacteria (42.6%), Acidobacteria (18.6%), Planctomycetes (9.0 %), Chloroflexi (Green nonsulfur bacteria, 7.5%), Bacteroidetes (2.1%), Gemmatimonadetes (2.7%), Nitrospirae (8.0%), Actinobacteria (High G+C Gram-positive bacteria, 6.4%) and candidate divisions (including the OP3, GN08, and SBR1093, 3.2%). Thirty-five clones were affiliated with bacteria that were related to nitrogen, sulfur, iron or manganese cycles. The comparison of the present data with the data obtained previously from caves based on 16S rRNA gene analysis revealed similarities in the bacterial community components, especially in the high abundance of Proteobacteria and Acidobacteria. Furthermore, this study provided the novel evidence for presence of Gemmatimonadetes, Nitrosomonadales, Oceanospirillales, and Rubrobacterales in a karstic hypogean environment.

• Horticultural Science & Technology
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• v.33 no.3
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• pp.396-402
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• 2015

This study identified a new male-sterile germplasm of Chinese jujube, named male-sterile No. 2 (JMS2), and achieved controlled hybridization using that germplasm as the female parent. The anthers of JMS2 before flower bud opening became shrunken, dingy yellow and much smaller than normal ones, and they changed to brown after anthesis. No pollen was observed in anthers of JMS2 and its male-sterile trait remained stable over different years. A total of 1,642 fruits were obtained from ten intra- and interspecific cross combinations via controlled hybridization from 2008 to 2012 using JMS2 as the female parent. Of those, 27.3% produced seeds, on average (0-72.6%). The rate of fruit with seed (RFS) was significantly different between cross combinations or years. Compared to other cross combinations, the RFS in the combination of JMS2 ${\times}$ 'Xingguang' (a Chinese jujube cultivar with high resistance to jujube witches' broom disease) and JMS2 ${\times}$ 'Xing16' (a sour jujube genotype) remained high in different years and reached means of 48.7 and 58.1%, respectively. Finally, 150 plantlets were regenerated from immature embryos, and 51 of them were randomly selected and identified to be authentic hybrids using amplified fragment length polymorphism (AFLP) markers. This is the first report of hybrids obtained from a cross between Chinese jujube and sour jujube.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.45 no.2
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• pp.123-127
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• 2000

The objective of this study was to develop a linkage map of soybean under the genetic background of Korean soybean. A set of 89 F/sub 5/ lines was developed from a cross between 'Pureunkong', which was released for soy-bean sprout, and 'Jinpumkong 2', which had no beany taste in seed due to lack of lipoxygenase 1, 2, and 3. A linkage map was constructed for this population with a set of 113 genetic markers including 7 restriction fragment length polymorphism (RFLP) markers, 79 randomly amplified polymorphic DNA (RAPD) markers, 24 simple sequence repeat(SSR) markers, and 3 morphological markers. The map defined approximately 807.4 cM of the soybean genome comprising 25 linkage groups with 98 polymorphic markers. Fifteen markers remained unlinked. Seventeen linkage groups identified here could be assigned to the respective 13 linkage groups in the USDA soybean genetic map. RFLP and SSR markers segregated at only single genetic loci. Fourteen of the 25 linkage groups contained at least one SSR marker locus. Map positions of most of the SSR loci and their linkages with RFLP markers were consistent with previous reports of the USDA soybean linkage groups. For RAPD, banding patterns of 13 decamer primers showed independent segregations at two or more marker loci for each primer. Only the segregation at op Y07 locus was expressed with codominant manner among all RAPD loci. As the soybean genetic map in our study is more updated, molecular approaches of agronomically important genes would be useful to improve Korean soybean improvement.

• The Plant Pathology Journal
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• v.25 no.1
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• pp.13-20
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• 2009

Ninety isolates of Colletotrichum species from new persimmon tree twigs and 50 isolates from pepper plant fruits were isolated via single-spore isolation. Of the 140 isolates, 26 were examined for mycelial growth, carbendazim sensitivity, and ITS sequence. Four of the isolates from the persimmon trees, which were cultivated exclusively in an orchard, showed fast mycelial growth and sensitivity to carbendazim, while five of the pepper isolates showed slower mycelial growth and were resistant to the fungicide. However, 17 isolates from persimmon trees cultivated with pepper plants in the same orchard showed slow mycelial growth like the pepper isolates and they were sensitive to carbendazim like the persimmon isolates. ITS sequence analysis of these 27 isolates led to the identification of the 22 persimmon isolates as C. gloeosporioides and the five pepper isolates as C. acutatum. PCR with species-specific primers confirmed that the 90 isolates from persimmon were C. gloeosporioides whereas the 50 isolates from pepper were C. acutatum. The 90 persimmon isolates of C. gloeosporioides and 50 pepper isolates of C. acutatum were compared by a wound inoculation test to determine their capacity for host cross-infection. All of the C. acutatum isolates from pepper caused typical symptoms of anthracnose on the fruits of pepper plants and twigs of persimmon; they differed from the C. gloeosporioides isolates from persimmon, more than 90% of which were able to infect only persimmon. Amplified fragment length polymorphism analysis revealed the existence of two groups (C. gloeosporioides and C. acutatum isolates group). At 80% genetic similarity, the C. gloeosporioides group was defined within four clusters, while the C. acutatum group was within three clusters. However, these clusterings were unrelated with the virulence of Colletotrichum species against pepper fruits.

• Journal of Plant Biotechnology
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• v.7 no.2
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• pp.81-86
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• 2005

Panax ginseng is one of the most important medicinal plants in the world. The international trade of ginseng is increasing yearly. The disguise of Chinese and American ginseng into Korean ginseng became a problem in recent years in abroad and Korea. An effective method to authenticate the Korean Panax ginseng from others at a DNA level is necessary for the healthy development of the ginseng market. Amplified fragment length polymorphism (AFLP) analysis was applied to develop a method for the identification of Korean ginseng between Chinese ginseng and American ginseng. It is very difficult to detect the different polymorphic bands among Korean field cultivated ginseng, and between field and wild-cultivated ginseng. The genetic distance coefficient by AFLP analysis between field- and wild cultivated Korean ginseng was very low, 0.056. Whereas, polymorphic bands between Korean and Chinese wild-cultivated ginseng was significantly different. The genetic distance coefficient between wild-cultivated Korean and Chinese ginseng was 0.149. The genetic distance coefficients between the P. ginseng and P. quinquefolius were ranging from 0.626 to 0.666. These results support that the AFLP analysis could be applied to authenticate Korean P. ginseng from others Chinese P. ginseng and American ginseng (P. quinquefolius).

• Parasites, Hosts and Diseases
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• v.35 no.4
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• pp.271-276
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• 1997

We tried to compare the three kinds oi Metagonimur species. M. Wokognulci, Ifetafonimus Miyata type, and M. tnknhashii, which were Know to be distributed in Korea with polymerase chain reaction based-restriction fragment length polymorphism (PCR- RFLP) patterns. We amplified the internal transcribed spacer 1 (ITSI) site of ribosomal RNA and mitochondrial cytochrome c oxidase I (mCOI) gene. The restriction patterns of ITSI gene loth Rsc I, ALu I and Msp I showed multiple fragmented bands of different sizes between three kinds of Metcgonimus. In case of mc01 gene, Rsc I and Alu I enzymes produced differentially fragmented band patterns. According to the parsimony analysis of PCR-RFLP patterns, the estimated genetic divergence between M Wokognwai and Metasoninus Miyata type was 0.034880, between Metusoninus Miyata type and M. tckc- hushii was 0.028098, between M. wokogawai and M. tnkahashii was 0.018179. It is suggested that Metasonimus Miyata type may be separate species and evolutionize at the older time than the other two species.

• Journal of Food Hygiene and Safety
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• v.15 no.2
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• pp.128-136
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• 2000

Twenty-four Vibrio strains were isolated from imported frozen seafoods and identified according to their physiological and biochemical properties. They included two V cholerae non-01 sp., two V. diazotrophicus sp., one V. hollisae sp., five V. natriegens sp., eight V. fluvialis sp., and four V. nereis sp.. Two of them were not identified as Vibrio species. When these strains were tested using API-2OE kit fur identification, however, only the results for two V. cholerae and five of the V. fluvialis strains matched the results obtained previously. Due to the importance of detecting V cholerae from foods, phylogenetic identification of the strains was attempted based on restriction fragment length polymorphism (RFLP) of the 16S rDNAs amplified by PCR. The results suggested that the two strains had identical RFLP patterns which were more closely related to that of V. proteolyticus than V. cholerae. The problems associated with identification of pathogens originated from seafoods demand development of accurate and rapid identification methods.

• Journal of Aquaculture
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• v.20 no.4
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• pp.255-259
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• 2007

Genetic diversity among four populations of Chinese shrimp Fenneropenaeus chinensis from Narodo, Yeonggwang, Taean and Chinese Bohai Bay was assessed by amplified fragment length polymorphism (AFLP) DNA fingerprinting. Total numbers of AFLP bands generated (ranging from 251 to 254) and average percent of polymorphic bands (27.1 to 28.1 %) were similar in the four populations. Heterozygosity and genetic diversity within or among the populations were very low for the populations with average values ranging from 0.1177 to 0.1288 and from 0.1099 to 0.1194, respectively. Analyses of pairwise distance, Fst index and genetic similarity among the populations also revealed the similar results with very low genetic differentiation each other. These results suggest that all the wild populations tested in the present analysis may be belonging to the same genetic origin, and also that they may have a close relationship in genetic structure without any significant differentiation.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.4
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• pp.564-570
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• 2016

The Sal-like 1 gene (Sall1) is essential for kidney development, and mutations in this gene result in abnormalities in the kidneys. Mice lacking Sall1 show agenesis or severe dysgenesis of the kidneys. In a recent study, blastocyst complementation was used to develop mice and pigs with exogenic organs. In the present study, transcription activator-like effector nuclease (TALEN)-mediated homologous recombination was used to produce Sall1-knockout porcine fibroblasts for developing knockout pigs. The vector targeting the Sall1 locus included a 5.5-kb 5' arm, 1.8-kb 3' arm, and a neomycin resistance gene as a positive selection marker. The knockout vector and TALEN were introduced into porcine fibroblasts by electroporation. Antibiotic selection was performed over 11 days by using $300{\mu}g/mL$ G418. DNA of cells from G418-resistant colonies was amplified using polymerase chain reaction (PCR) to confirm the presence of fragments corresponding to the 3' and 5' arms of Sall1. Further, mono- and bi-allelic knockout cells were isolated and analyzed using PCR-restriction fragment length polymorphism. The results of our study indicated that TALEN-mediated homologous recombination induced bi-allelic knockout of the endogenous gene.

• Journal of Oral Medicine and Pain
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• v.21 no.2
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• pp.351-367
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• 1996

In the field Of individual identification in forensic Science, if the body is charred, it is sometimes impossible to identify the morphologic changes and charred tissue such as blood, muscle and bone can not be identified by forensic microbiologic method such as DNA typing. So the author used the characteristics of teeth which is relatively firm compare to other organs and stable to external environment such as heat and also possess cells needed for the DNA typing. The author conducted the experiment on teeth to detect DNA related to individual identification regarding to temperature in which other charredorgans can not be detected. The experiment was done on 64 extracted third molars consisted of unheated ones, and heated teeth to $100^{\circ}C$, $150^{\circ}C$, $200^{\circ}C$ for 45 min, 90 min, and 120 min respectively and to $250^{\circ}C$ for 45 min. DNA was extracted from each tooth and amplified fragment length polymorphism procedure(AMP-FLPs) using polymerase chain reaction(PCR) was applied and observed for the matching DNA in HumTH01 and HumCD4 locus and the followings Are the results : 1. It was able to detect matching DNA in HumTH01 and HumCD4 locus in every teeth which no heating has been done. 2. It was able to detect matching DNA in HumTH01 and HumCD4 locus in every teeth heated to $100^{\circ}C$ for 45, 90 and 120 min. 3. It was able to detect matching DNA in HumTH01 and HumCD4 locus in teeth heated to $l00^{\circ}C$, $200^{\circ}C$ for 45, 90, 120 min. 4. It was impossible to detect matching DNA in HumTH01 and HumCD4 locus in teeth heated to $250^{\circ}C$. So, it is possible to extract DNA from teeth that otherwise can not be extracted from other organs in the charred body and it can be concluded that teeth are highly reliable and applicatable as forensic odontology for individual identification.