• Korean Journal of Breeding Science
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• v.41 no.4
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• pp.420-428
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• 2009

In this study, the variation of 20 Pinus thunbergii isolates of F. circinatum from 2 damaged sites in Jeju-Island were compared with a known Fusarium circinatum using molecular biological techniques. Two- and four-year-old seedlings of Pinus rigida${\times}$Pinus x rigitaeda and two-, three- and six-year-old seedlings of P. thunbergii were inoculated with one of the most virulent isolates, FT-7, to determine differences in susceptibility. In site 1 (FT), 13 isolates of F. circinatum were isolated from 14 individuals and in site 2 (FS), 7 isolates of F. circinatum were isolated from 9 individuals. No difference was found in the sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA genes in the FS and FT isolates, and also even in the known isolate of F. circinatum, FE 1-1. However, the ITS sequences of the FS and FT isolates differed from those of a fungus, Botrytis cinerea. Two-year-old seedlings of P. rigida${\times}$P. x rigitaeda showed higher susceptibility (93.3% of mortality) than four-year-old ones. Three-year-old seedlings of P. thunbergii showed the highest susceptibility (66.7% of mortality) compared to those at other ages in the same species. We found a positive correlation between basal diameter and lesion length in the seedlings of P. rigida${\times}$P. x rigitaeda ($R^2=0.66$) and P. thunbergii (p < 0.0001), respectively. There were significant differences in susceptibility by the age of seedlings in each of P. rigida${\times}$P. x rigitaeda (p < 0.0001) and P. thunbergii (p < 0.0001) based on lesion length.

• Applied Biological Chemistry
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• v.15 no.2
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• pp.93-109
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• 1972

Three lots of Chung-Kook-Jang were prepared by the use of 2 strains of Bacillus subtilis and Bacillus natto. For four samples taken from each lot in 12 hrs interval changes of nitrogenous compounds, insoluble protein, water soluble protein, peptides, free amino acids, amino and ammonia nitrogens during Chung-Kook-Jang fermentation, were studied together with the changes of moisture, pH, proteolytic enzyme activity. In addition the average peptide length of the peptides of a Bacillus subtilis lot was determined by the method of molecular sieving using ion exchange resin. The results were as follows: 1. The contents of moisture and total-nitrogen changed little in all samples throughout the fermentation as it would be expected. 2. In all three experimental lots the pH became higher gradually from the initial value of 6.65 to the final $7.5{\sim}7.85$ during the fermentation. Proteolytic enzyme activities, in accordance with this pH change, steadily increased up to $48{\sim}60$ hrs. of fermentation and then slightly decreased, probably affected by the high pH. The most strong proteolytic activity was observed in the experimental Chung-Kook-Jang fermentation lot using the Bacillus subtilis K-27 isolated by the author. 3. The contents of insoluble protein nitrogen in soybeans increased markedly (5%) by the cooking, after steeping 12 hrs in water. During the Chung-Kook-Jang fermentation, however, it decreased from 1/2 to 1/10 of that of the cooked soybeans. 4. The contents of water soluble protein nitrogen (5%) whereas, greatly decreased to the value of 1.0% by the cooking; but little changed further during the fermentation, 5. The total contents (0.25%) of peptides, amino, and ammonia-nitrogens, PAA-N., increased almost double by the cooking and steadily became higher as the fermentation proceeded, reaching finally up to$4{\sim}7%$ in 72 hrs fermentation. 6. The amounts of free amino acids of soybean generally decreased during the processing of cooking, even some of them like glutamic acid were destroyed completely, However in the subsequent Chung-Kook-Jang fermentation for 72 hrs., they showed from several to a few hundreds folds increases depending upon the kinds of amino acids. Valine which was contained in HCl-hydrolyzed steeped or cooked soybeans in amounts $220{\sim}267mg%$ was not detected at all as the free amino acid in all fermented samples. 7. Average peptide length (APL) of all fractions, eluted and fractionated by using the Dowex-50 ion exchange resin column, and fraction collector showed the highest value for the cooked soybean and then decreased as the fermentation proceeded. The APL value of effluent showed the highest in 12 hrs fermented sample, The value decreased thereafter by fermentation.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.2
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• pp.99-110
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• 2008

Objectives: Preimplantation genetic diagnosis (PGD) has become an assisted reproductive technique for couples carrying genetic conditions that may affect their offspring. Osteogenesis imperfecta (OI) is an autosomal dominant disorder of connective tissue characterized by bone fragility and low bone mass. At least 95% of cases are caused by dominant mutations in the COL1A1 or COL1A2. In this study, we report on our experience clinical outcomes with 5 PGD cycles for OI in two couples. Methods: Before clinical PGD, we assessed the amplification rate and allele drop-out (ADO) rate of alkaline lysis and nested PCR protocol using heterozygous patient's single lymphocytes in the pre-clinical diagnostic tests for OI. We performed 5 cycles of PGD for OI by nested PCR for the causative mutation loci, COL1A1 c.2452G>A and c.3226G>A, in case 1 and case 2, respectively. The PCR products were analyzed by agarose gel electrophoresis, restriction fragment length polymorphism (RFLP) analysis with HaeIII restriction enzyme in the case 1 and direct DNA sequencing. Results: We confirmed the causative mutation loci, COL1A1 c.2452G>A in case 1 and c.3226G>A in case 2. In the pre-clinical tests, the amplification rate was 94.2% and ADO rate was 22.5% in case 1, while 98.1% and 1.9% in case 2, respectively. In case 1, a total of 34 embryos were analyzed and 31 embryos (91.2%) were successfully diagnosed in 3 PGD cycles. Eight out of 19 embryos diagnosed as unaffected embryos were transferred in all 3 cycles, and in the third cycle, pregnancy was achieved and a healthy baby was delivered without any complications in July, 2005. In case 2, all 19 embryos (100.0%) were successfully diagnosed and 4 out of 11 unaffected embryos were transferred in 2 cycles. Pregnancy was achieved in the second cycle and the healthy baby was delivered in March, 2008. The causative locus was confirmed as a normal by amniocentesis and postnatal diagnosis. Conclusions: To our knowledge, these two cases are the first successful PGD for OI in Korea. Our experience provides a further demonstration that PGD is a reliable and effective clinical techniques and a useful option for many couples with a high risk of transmitting a genetic disease.

• Tuberculosis and Respiratory Diseases
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• v.63 no.3
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• pp.235-241
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• 2007

Background: Airway mucus hypersecretion plays an important role in the pathogenesis of asthma, and is associated with the induction of MUC5AC expression in airway secretion. The MUC5AC gene is highly polymorphic; however, there are few studies about the association between the polymorphisms of the MUC5AC gene and asthma susceptibility or asthma phenotypes. We have investigated the association of MUC5AC promoter polymorphisms with the risk of asthma and asthma phenotypes. Methods: We determined the genotypes of the MUC5AC promoter (-1274G>A) in 78 asthma patients and in 78 age, sex-matched control individuals in the Korean population. Genomic DNAs from blood were analyzed by PCR and RFLP (restriction fragment length polymorphism) determination. We examined $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy (by a skin test) in asthma patients. Results: The mean age of the patients was $47.7{\pm}16.1$ years and 38.5% were men, and the mean $FEV_1$ was $84.4{\pm}22.3%$ of predicted in the asthma patients. The -1274G>A polymorphism of the MUC5AC promoter in asthma patients was not significantly different as compared with normal individuals (GG 57.7%, AG 34.6% and AA 7.7% in asthma patients vs. GG 56.4%, AG 38.5% and AA 5.1% in control subject, p = 0.752, Cod). Several clinical parameters in asthma patients such as $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy, were not associated with the -1274G>A polymorphism of the MUC5AC promoter. Conclusion: The -1274G>A single nucleotide polymorphism (SNP) frequency of the MUC5AC promoter was not associated with asthma in a Korean population.

• Childhood Kidney Diseases
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• v.3 no.2
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• pp.209-216
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• 1999

Purpose : Nephrogenic diabetes insipidus (NDI) is a rare X-linked disorder associated with renal tubule resistance to arginine vasopressin (AVP). The hypothesis that the defect underlying NDI might be a dysfunctional renal AVPR2 has recently been proven by the identification of mutations in the AVPR2 gene in NDT patients. To investigate the association of mutations in th AVPR2 gene with NDI, we analyzed the AVPR2 gene located on the X chromosome. Methods : We have analyzed the AVPR2 gene in a kindred with X-linked NDI. The proband and proband's mother were analyzed by polymerase chain reaction-single strand conformational polymorphism(PCR-SSCP) and DNA sequencing of the AVPR2 gene. We also have used restriction enzyme analysis of genomic PCR product to evaluate the AVPR2 gene. Results : C to T transition at codon 202, predictive of an exchange of tryptophan 202 by cysteine(R202C) in the third extracellular domain was identified. This mutation causes a loss of Hae III site within the gene. Conclusion : We found a R202C missense mutation in the AVPR2 gene causing X-linked NDI, and now direct mutational analysis is available for carrier screening and early diagnosis.

• Korean Journal of Weed Science
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• v.31 no.2
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• pp.134-145
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• 2011

The objectives of this study were to investigate fitness difference in growth and rice yield in herbicide-transgenic rice overexpressing Myxococcus xanthus and Arabidopsis thaliana protoporphyrinogen oxidase (Protox) genes and non-transgenic rice. We also aimed to determine whether these fitness differences are related to ALA synthesizing capacity, accumulation of terapyrroles, reactive oxygen species, lipid peroxidation, and antioxidative enzymes at different growth stages of rice. Plant height of the transgenic rice overexpressing M. xanthus (MX) and A. thaliana (AP37) Protox genes at 43, 50, and 65 days after transplanting (DAT) was significantly lower than that of WT. Number of tiller of PX as well as MX and AP37 at 50 and 65 DAT was significantly lower than that of WT. At harvest time, culm length and yield of MX, PX and AP37 and rice straw weight of MX and AP37 were significantly low compared with WT. The reduction of yield in MX, PX, and AP37 was caused by spikelets per panicle and 1000 grain weight, ripened grain, spikelets per panicle, 1000 grain weight, and ripened grain, respectively. On the other hand, 135 the reduction of yield in MX, PX, and AP37 was also observed in another yearly variation experiment. The reduction of rice growth in MX, PX, and AP37 was observed in seedling stage as well as growth duration in field. There were no differences in tetrapyrrole intermediate Proto IX, Mg-Proto IX and Mg-Proto IX monomethyl ester, reactive oxygen species ($H_2O_2$ and ${O_2}^-$), MDA, antioxidative enzymes (SOD, CAT, POX, APX, and GR) and chlorophyll between transgenic lines and wild type, indicating that accumulated tetrapyrrole intermediate and other parameters were not related to growth reduction in transgenic rice. However, ALA synthesizing capacity in MX, PX, and AP37 at one day after exposure to light and 52 DAT was significantly lower than that of WT. Further study is required to elucidate the mechanisms underlying the growth and yield difference between transgenic and WT lines.

• Journal of Sericultural and Entomological Science
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• v.51 no.2
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• pp.211-217
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• 2013

In general, the mean silkmoth lifespan is around 8 days for female and 5 days for male. But, the duration of J037 strain's lifespan is remarkably long in both sexes. On the contrary, the Daizo(sdi) strain has a remarkably short lifetime. The differences in adult lifetime among various silkworm strains has been suggested that the adult lifetime may be genetically controlled. In this experiment, using J037 and Daizo strains we investigated genetic factors related to the adult lifetime of silkworm. We constructed the full-length cDNA library from the adult male of the J037 strain. A total of 2,688 clones were randomly selected, and we performed a differential display hybridization with cDNA probes generated from J037 and Daizo adult males. In conclusion, 193 clones were identified as differential expressed genes, and 154 unique genes were generated after the assembly of 193 clones. Of the 154 unique genes, the most abundant genes were cytochrome oxidase subunit-1 gene(9 times) and unknown(clone ID; 1-50) gene(5 times). The functional groups of these unique genes with matches in the AmiGo database were constructed according to their putative molecular functions. Among thirteen functional categories, the largest group was unclassified protein(24%). In addition, we analyzed the nucleotide and deduced amino acid sequences of the most highly occurred gene(1-50, EF434397), which consisted of 240 amino acids. However, it is confirmed yet that these genes really have an affected on the silkworms longevity. Further studies on these molecules biological roles will give us well-fined information about mechanisms of insect aging and/or scenesence.

• Journal of the korean academy of Pediatric Dentistry
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• v.32 no.1
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• pp.75-88
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• 2005

Ameloblastoma is the most commonly occurring odontogenic tumor in oral cavity. Although most are benign epithelial neoplasm, they are generally considered to be locally aggressive and destructive, exhibiting a high rate of recurrence. The biological behavior of this neoplasm is a slowly growing, locally invasive tumor without metastasis, therefore malignant neoplasm, changed its histological appearance to carcinoma or showed distant metastasis, is only defined clinically. In this study, we identified the differentially expressed genes(DEGs) in stages under benign or malignant ameloblastoma compared with normal patient using ordered differential display(ODD) reverse transcription(RT)-PCR and $GeneFishing^{TM}$ technology. ODD RT-PCR is rather effective when the investigation of samples containing very small amounts of total RNA must be accomplished. ODD RT-PCR used the means of amplification with anchored T-primer and adaptor specific primer. bearing definite two bases at their 3' ends and so this method could display differential 3'-expressed sequence taqs(ESTs) patterns without using full-length cDNAs. Compared with standard differential display, ODD RT-PCR is more simple and have enough sensitivity to search for molecular markers by comparing gene expression profiles, However, this method required much effort and skill to perform. $GeneFishing^{TM}$ modified from DD-PCR is an improved method for detecting differentially expressed genes in two or more related samples. This two step RT-PCR method uses a constant reverse primer(anchor ACP-T) to prime the RT reaction and arbitrary primer pairs(annealing control primers, ACPs) during PCR. Because of high annealing specificity of ACPs than ODD RT-PCR, the application of $GeneFishing^{TM}$ to DEG discovery generates reproducible, authentic, and long(100bp to 2kb) PCR products that are detectable on agarose gels. Consequently, various DEGs observed differential expression levels on agarose gels were isolated from normal, benign, and malignant tissues using these methods. The expression patterns of the some isolated DEGs through ODD RT-PCR and $GeneFishing^{TM}$ were confirmed by Northern blot analysis and RT-PCR. The results showed that these identified DEGs were implicated in ameloblastoma neoplasm processes. Therefore, the identified DEGs will be further studied in order to be applied in candidate selection for marker as an early diagnosis during ameloblastoma neoplasm processes.

• Journal of Plant Biotechnology
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• v.32 no.3
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• pp.167-173
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• 2005

Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 1995.06b
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• pp.77-96
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• 1995

To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.