• Journal of Chest Surgery
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• v.30 no.9
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• pp.854-861
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• 1997

CYPRA 21-1 is known to be a cytokeratin 19 fragment, and it can be detected by using two specific monoclonal antibodies (KS 19-1 and BM 19-21) and can be clinically applied as a useful circulating tumor marker The epidermal growth factor receptor (EGF-R) expression was evaluated and characterized by its tyrosine protein kinase activity and by its ligand-stimulated autophosphorylation, a property shared with other peptide growth factor receptors. Autocrine or para'urine action was initiated by a growth factor, or by a transforming growth factor o, which had an extensive homology with EGP and which also stimulated tyrosine kinase activity on the EGF-R. The CYFRA 21-1 and the EGF-R levels in 30 patients with primary lung tumors were investigated. There were 24 patients with squamous cell carcinomas and 6 patients with adenocarcinomas. Specimen 5 mm3 in size were sampled at three different locations ; the main lesion, the boundary between the lesion and the unaffected tissue, and the unaffected tissue of the patients. The results were as follows 1. The CYPRA 21-1 concentration in the cancer boundary, the most malignant region,(348.6 : 89.9 ng/ml) was the lowest value. The CYFRA 21-1 concentration in unaffected tissue,(718.4$\pm$77.8 ng/ml) was higher than that in the main lesion. which had intact cellularity. 2. The EGF-R concentration in the main lesion was higher than that in the unaffected tissue, and the EGF-R concentration in a squamous cell cacinoma was higher than that in an adenocarcinoma. also, the EGF-R concentration in the cancer b undary was highest at stage 1, ll. The EGF-R concentration was higher in the main cancer lesion that in the unaffected tissue at stage 111, IV. 3. The CYFRA 21-1 was a cytoplasmic skeleton and the EGF-R was a cell-wall component; there was no correlation. In conclusion, CYFRA 21-1 was abundant in the cytoplasm but had a higher concentration in the unaffected tissue than in the main cancer lesion. The CYFRA 21-1 concentration of the tissue did not reflect the amount of cancer activity, the EGP-R was located in the cell membrane, the level of tissue that reflects cancer activity, so the main cancer lesion had a higher concentration than the unaffected tissue. CYFRA 21-1 is not a useful tumor maker at the tissue level. Because the EGF-R concentration re(looted the cancer activity, its a useful tumor marker for lung cancer.

• Korean Journal of Environment and Ecology
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• v.22 no.3
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• pp.309-319
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• 2008

Roads are an indicator of anthropogenic activity causing ecosystem disturbances and often lead to habitat fragmentation, habitat loss, and habitat isolation. The Hallasan National Park(153.4$km^2$) on Jeju Island being distinguished for its unique geology, topography, and biota has also been designated as a core area of UNESCO Man and the Biosphere(MAB) Reserve. Although the high conservation value of this park has contributed to a rapid growth of tourists and road construction, landscape changes due to roads have not been examined yet. We used GIS systems to examine the fragmentation pattern caused by roads, in relation to its zonation, elevation, and vegetation. When a buffer was applied to roads(112m width for paved roads and 60m width for both legal and illegal trails), the park consisted of 100 fragments. The ten fragments generated after applying buffer to only paved roads and legal trails ranged from $0.002km^2$ to $38.2km^2$ with a mean of $14.2km^2$, and about 7% of both nature conservation zone and nature environment zone of the park were edge. Fragments in both east and west ends of the park and around the summit exhibited relatively high shape indices with means of 5.19(for 100 fragments) and 7.22(for 10 fragments). All five legal trails are connected to the pit crater of the mountain and vegetation changed from broadleaf forests and conifer forests to grasslands with elevation, consequently resulting in dramatic fragment size reduction in grasslands at high elevation, in particular above 1,400m, where endemic and alpine plants are abundant. These results show that in Hallasan National Park the risks of habitat deterioration and habitat loss due to fragmentation may be more severe in the nature conservation zone dominated by Baengnokdam than in the nature environment zone. Therefore, current road networks of the park appear to fall short of the goal of the national park for ecosystem conservation and protection. Considering that the entire Hallasan National Park also serves as a MAB core area, conservation efforts should focus, first of all, on park rezoning and road management to mitigate habitat fragmentation.

• Korean Journal of Ichthyology
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• v.18 no.4
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• pp.300-310
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• 2006

Genomic DNA isolated from three geographical gizzard-shad (Konosirus punctatus) populations in Seocheon (SC), Busan (BS) and Gochang (GC) collected in the West Sea and the southern sea, respectively, off the Korean Peninsula, were PCR-amplified repeatedly. Eight selected decamer and 20-mer primers generated a total of 713 loci in the SC population, 791 in the BS population, and 732 in the GC population, with a DNA fragment size ranging from 100 bp to 2,800 bp. We identified 50 unique loci for the SC population, 70 unique loci for the BS population and 130 for the GC population: 120 shared loci for the three populations. There were 108 specific loci (15.1%) for the SC population, 74 (9.4%) for the BS population, and 67 (9.2%) for the GC population. Eight primers also generated 48 polymorphic loci (6.7%) for the SC population, 26 (3.3%) for the BS population, and 16 (2.2%) for the GC population. The similarity matrix ranged from 0.756 to 0.936 for the SC population, from 0.800 to 0.938 for the BS population, and from 0.731 to 0.959 for the GC population. The dendrogram obtained by the eight primers indicates three genetic clusters: cluster 1 (SEOCHEON 01~SEOCHEON 10), cluster 2 (BUSAN 11~BUSAN 20 and GOCHANG 23~GOCHANG 24), and cluster 3 (GOCHANG 21, 22, 25, 26, 27, 28, 29 and 30). As stated above, some individuals of the GC population appear to belong in BS population. When seeing this result, it was thought with the fact that some individuals of 2 populations seem to come and go partially. Thus, RAPD-PCR analysis revealed a significant genetic distance between the three geographical gizzard-shad populations. Using various decamer and 20-mer primers, RAPD-PCR may be applied to identify specific/polymorphic markers that are particular to a species and geographic population, and to define genetic diversity, polymorphisms, and similarities among geographical gizzard-shad populations.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.12
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• pp.575-580
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• 2017

This study analyzed the effects of RGG box of hnRNP A1 on its subcellular localization and stabilization of hnRNP A1 over a three year period from October 2014. First, a 6R/K mutation in RGG box was generated, and pcDNA1-HA-hnRNP A1(6R/K) was constructed. The subcellular localization of hnRNP A1(6R/K) from the HeLa cells transfected with this plasmid DNA was analyzed by immunofluorescence microscopy. HA-hnRNP A1(6R/K) was found to exhibit nuclear and cytoplasmic fluorescence. The stability of hnRNP A1(6R/K) was checked by Western blot analysis using the expressed protein from the HeLa cells transfected with the pcDNA1-HA-hnRNP A1(6R/K). The results show that HA-hnRNP A1(6R/K) has a smaller size. These confirm that HA-hnRNP A1(6R/K) is localized both in the nuclear and cytoplasm, not because 6R/K mutation affects the nuclear localization of hnRNP A1, but because 6R/K mutation causes hnRNP A1(6R/K) to cleave at the mutation or near the mutation site. The cleaved protein fragment, which lacks the M9 domain (i.e. nuclear localization signal of hnRNP A1), did not exhibit nuclear fluorescence. This suggests that the arginines of RGG box in hnRNP A1 play an important role in stabilizing hnRNP A1. An analysis of the RNA-binding ability of hnRNP A1(6R/K) expressed and purified from bacteria will be a subsequent research project.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.17-25
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• 2013

The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag, $2{\times}(Gly_4Ser_1)$ linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.

• Horticultural Science & Technology
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• v.18 no.3
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• pp.342-347
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• 2000

The objectives of this study were to investigate the genetic and phenotypic features of male sterile transformants by pollen-specific expression of diphtheria toxin gene and to find out inheritance patterns of transgene to the next generation. When backcrossed (BC) progenies were tested for expression of kanamycin resistance ($Km^R$), 9 lines out of 13 lines, except 4 lines ($BC_{1}5-13,\;BC_{1}5-23,\;BC_{1}5-28,\;BC_{1}5-32$), showed the ratio of $Km^R$ to kanamycin sensitive ($Km^S$), from 1:30 to all $Km^S$. As a result, they were much lower than Mendelian segregation of a dominant gene. To determine whether male sterility is a heritable and stable trait, 5 male sterile plants ($BC_{1}5-13,\;BC_{1}5-14,\;BC_{1}5-23,\;BC_{1}5-32,\;BC_{1}5-33$ lines) which had different transgene copy numbers were backcrossed as female parents with pollens from wild type. To confirm the existence of the DTx-A gene in the genome of the progenies, PCR was conducted using specific primers of the DTx-A coding region. A PCR band of 428 bp was obtained from each generation, which is the predicted size of the DTx-A gene fragment. Trangenes were inherited to the next $BC_4T_0$ progenies and showed male sterility, however, based on the copy numbers of DTx-A gene male sterile plants did not show predicted ratio. When male sterile plants were backcrossed with fertile plants, fruit capsule sizes and seed settings were relatively reduced from those of selfing wild type plants. The fruit sizes and seed settings were reduced in proportion to the increase in the copy number of DTx-A gene.

• Research in Plant Disease
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• v.15 no.2
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• pp.83-87
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• 2009

Direct Polymerase Chain Reaction (PCR) method that combines biological and enzymatic amplification of PCR targets was developed for the detection of Xanthomonas axonopodis pv. glycines on soybeen seeds without DNA isolation. Primers Xag F1 and Xag R1 were designed to specifically amplify a 401 bp fragment of the glycinecin A gene of X axonopodis pv. glycines. Xag F1 and Xag R1 were used to carry out the PCR analysis with genomic DNA from 45 different bacterial strains including phylogenetically related bacteria with X axonopodis pv. glycines, and other bacterial strains of different genus and species. The PCR assay using this set of primers were able to detect X axonopodis pv. glycines with DNA concentration as low as 200 fg and $1.8{\times}10^3$ cfu/ml. The Xag was detected from the seed samples incubated for 2 hrs with shaking and the intensity of the band was increase with the incubation time of seeds. The Direct PCR assay method without DNA isolation makes detection of X. axonopodis pv. glycines on soybean seeds easier and more sensitive than other conventional methods. The developed seed assay using direct PCR method will be useful for the specific detection of X. axonopodis pv. glycines in soybean seed samples.

• Journal of Animal Science and Technology
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• v.48 no.5
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• pp.637-650
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• 2006

Genomic DNA isolated from two crucian carp species obtained from Yesan (Carassius auratus) and Dangjin (Carassius cuvieri) in Korea were amplified at several times by polymerase chain reaction (PCR). The amplified products were separated by agarose gel electrophoresis (AGE) with oligonucleotides decamer primer and stained with ethidium bromide. The seven arbitrarily selected primers OPC-11, OPC-14, OPC-18, OPD- 02, OPD-11, OPD-15 and OPD-20 generated the shared loci by each species, the polymorphic and specific loci. The seven primers generated the total 458 loci that can be scored from the crucian carp obtained in C. auratus species. 358 fragments were generated from the species obtained in C. cuvieri species. The size of DNA fragments varies from 150 to 1,600bp. The complexity of the banding patterns varies dramatically between the primers and two locations. In this study, 458 loci were identified in the crucian carp species from Yesan and 358 in the crucian carp species from Dangjin: 84 polymorphic loci (18.3%) in the C. auratus species and 48 (13.4%) in the C. cuvieri species. 154 shared loci by each species, the average 22 per primer, were observed in the C. auratus species and 187 loci, the average 26.7 per primer, in the Dangjin species. Based on the average bandsharing (BS) values of all samples, the similarity matrix ranged from 0.434 to 0.868 in the C. auratus species and from 0.449 to 0.924 in the C. cuvieri species. The average BS value was 0.641±0.013 within the C. auratus species and 0.684±0.013 within the C. cuvieri species. The average BS value between two crucian carp species 0.484 ± 0.007, ranged from 0.307 to 0.682. The BS value between the individual No. 09 and No. 16 was 0.682, which was the highest between two crucian carp species. Compared separately, the BS value of individuals within the C. cuvieri species was higher than the C. auratus species. The dendrogram obtained by the seven primers, indicates three genetic clusters: cluster 1 (AURATUS No. 01, 02, 03, 04, 05, 06, 07, 08, 09, 10 and 11), cluster 2 (CUVIERI No. 12, 13, 14, 15, 16, 17, 18, 19, 20 and 21) and cluster 3 (CUVIERI no. 22). The shortest genetic distance displaying significant molecular difference was between the individual AURATUS No. 09 and AURATUS No. 08 from Yesan (genetic distance=0.064). The longest genetic distance displaying significant molecular differences was between the individual CUVIERI No. 17 and AURATUS No. 11 between two crucian carp species (0.477). RAPD-PCR analysis has revealed the significant genetic distance between two crucian carp species pairs.(Key words: Carassius auratus, Carassius cuvieri, Crucian Carp, DNA Polymorphism, Genetic Distance)

• Horticultural Science & Technology
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• v.28 no.5
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• pp.828-835
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• 2010

Conventional methods for identification of apple cultivars are based on the evaluation of sets of morphological characteristics, however, closely related cultivars often cannot be distinguished by morphological traits. This study was conducted to develop DNA markers for discrimination of the apple cultivars bred in Korea. Thirty random primers generated eighty-three random amplified polymorphic DNA (RAPD) markers from thirty-one Korean bred and introduced apple cultivars. Fifty-two RAPD fragments were cloned and sequenced for conversion into sequence characterized amplified region (SCAR) markers. Among them only seventeen SCAR markers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Several combinations of six (AN11_433, AN08_566, A408_592, AK17_653, AO04_711, AO04_779 or AW15_368, AN11_433, A408_592, AK17_653, AO04_711, AO04_779, or AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AO04_779) to seven (AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AM16_708, AO04_779 or A330_424, AN11_433, AG14_502, AN08_566, A408_592, AK17_653, AO04_779 or A330_424, AN11_433, AK14_564, A408_592, AK17_653, AM16_708, AT14_789) SCAR markers provided enough polymorphism to identify sixteen Korean apple cultivars among thirty-one tested cultivars. Therefore, application of the seventeen SCAR markers was sufficient to identify the thirty-one tested apple cultivars. These markers could be utilized as a reliable tool for cultivar discrimination of Korean apples.

• Journal of the Korean Wood Science and Technology
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• v.37 no.3
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• pp.255-264
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• 2009

To evaluate the effects of chemical pretreatments of lignocellulosic biomass on enzymatic hydrolysis process, Populus euramericana was pretreated for 1 hr with 1% sulfuric acid ($H_2SO_4$) at $150^{\circ}C$ and 1% sodium hydroxide (NaOH) at $160^{\circ}C$, respectively. Before the enzymatic hydrolysis, each pretreated sample was subjected to drying process and thus finally divided into four subgroups; dried or non-dried acid pretreated samples and dried or non-dried alkali pretreated samples and chemical and physical properties of them were analyzed. Biomass degradation by acid pretreatment was determined to 6% higher compared to alkali pretreatment. By the action of acid ca. 24.5% of biomass was dissolved into solution, while alkali degraded ca. 18.6% of biomass. However, reverse results were observed in delignification rates, in which alkali pretreatment released 2% more lignin fragment from biomass to the solution than acid pretreatment. Unexpectedly, samples after both pretreatments were determined to somewhat higher crystallinity than untreated samples. This result may be explained by selective disrupture of amorphous region in cellulose during pretreatments, thus the cellulose crystallinity seems to be accumulated in the pretreated samples. SEM images revealed that pretreated samples showed relative rough and partly cracked surfaces due to the decomposition of components, but the image of acid pretreated samples which were dried was similar to that of the control. In pore size distribution, dried acid pretreated samples were similar to the control, while that in alkali pretreated samples was gradually increased as pore diameter increased. The pore volume which increased by acid pretreatment rapidly decreased by drying process. Alkali pretreatment was much more effective on enzymatic digestibility than acid pretreatment. The sample after alkali pretreatment was enzymatically hydrolyzed up to 45.8%, while only 26.9% of acid pretreated sample was digested at the same condition. The high digestibility of the sample was also influenced to the yields of monomeric sugars during enzymatic hydrolysis. In addition, drying process of pretreated samples affected detrimentally not only to digestibility but also to the yields of monomeric sugars.