Differentially Expressed Gene (DEG) was obtained from Differential Display Reverse Transcription (DDRT)-PCR using Annealing Control Primer (ACP) to search and clone genes related to developmental stages of Sebastes inermis. By using 120 ACPs, the nucleotide sequences obtained from 16 DEGs showing higher expression in 6-month-old skeletal muscle than 18-month-old ones and from 22 DEGs displaying stronger expression in 18-month-old than 6-month-old were analyzed and BLAST was conducted. The results identified that DEGs shared 69~95% homology with genes of parvalbumin (PVALB), nucleoside diphosphate kinase (NDK) B, tropomyosin (TPM), troponin I (TnI), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), muscle-type creatine kinase (CKM2), small EDRK-rich factor 2 (SERF2), adenosine monophosphate deaminase (AMPD), Trimeric intracellular cation channel type A (TRICA), Rho GTPase-activating protein 15 (ARHGAP15), S-formylglutathione hydrolase (Esterase D; ESD), heat shock protein 70 (hsp70), type 1 collagen alpha 2 (COL1A2), glutathione S-transferase, Mid1-interacting protein 1 (Mid1lip1), myosin light chain 1 (MYL1), sarcoplasmic/endoplasmic reticulum calcium ATPase 1B (SERCA1B), and ferritin heavy subunit (FTH1). Expression pattern by developmental stage of DEG14 and PVALB exhibiting strong expression in 6-month-old skeletal muscle was investigated using real time PCR. Expression was reduced as Sebastes inermis grew. Expression of PVALB gene was extremely low after 6 months of age. Expression of CKM2 showed higher expression in 18-month-old skeletal muscle than in 6-month-old muscles, and increased continuously until 4 years old, after which CKM2 expression became gradually reduced. By analysis of tissue-specific expression patterns of DEG, DEG14 was expressed mainly in skeletal muscle, liver, kidney and spleen tissues, whereas PVALB expression was expressed in skeletal muscle and kidney, but not in liver and spleen tissues. CKM2 was expressed in skeletal muscle, kidney, and spleen tissues, but not in liver tissues. PVALB gene was composed of 110 amino acids, which constituted 659 bp nucleotides. The results reported here demonstrate that the expression patterns of parvalbumin and CKM2 could be used as molecular markers for selecting fishes exhibiting fast growth.
Pisidium (Neopisidium) coreanum is a freshwater snail and lives in spring water near mountain areas. Interestingly, this snail has been traditionally regarded as medicinal food, and thus has been used as folk remedies for healing broken bones. Recently, alpha classification on Pisidium (Neopisidium) coreanum through redescription has been conducted. However, not much attention has been made in beta classification. In this study, we performed the beta classification based on metallothionein (MT) genes found from various organisms. To this end, the complete cDNA sequences were obtained from the Expressed Sequence Tag (EST) sequencing project of Pisidium (Neopisidium) coreanum. The coding region (315 bp) encoded an amino acid sequence of 105 residues. The combined results from BLAST analyses, multiple sequence alignment and molecular phylogenetic study of Pc-MT gene indicate that Pisidium (Neopisidium) coreanum has similarity to freshwater bivalve such as Dreissena polymorpha (zebra mussel), Unio tumidus (swollen river mussel) and Crassostrea ariakensis (suminoe oyster).
Park, So Young;Jeong, Ji Eun;Hwang, Hee Ju;Wang, Tae Hun;Park, Eun Bi;Kim, Yong Min;Lee, Jun-Sang;Han, Yeon Soo;Yang, Seung-Ha;Lee, Yong Seok
The Korean Journal of Malacology
/
v.30
no.2
/
pp.155-163
/
2014
Serpins are a group of proteins involved in the regulation of serine and other type of proteases, and have been identified in many kinds of organisms from invertebrates to vertebrates. Serpins are known to regulate the proteolytic cascades of the innate immune pathways in addition to their roles in blood coagulation, angiogenesis, fibrinolysis, inflammation and tumor suppression. In this study, we have isolated two partial serpin gene fragments from expressed sequence tags (ESTs) of Nesiohelix samarangae. Dotplot analysis indicates that they are of two different types, Ns-serpin type 1 and Ns-serpin type 2. Ns-serpin type 1 has 819 bp coding region (272 amino acids), whereas Ns-serpin type 2 has 555 bp coding region (185 amino acids). Molecular phylogenetic analysis shows that the identified serpins have high similarities to their counterparts in the California see slug, Aplysia californica. Yet, the precise biological and immunological roles of these Ns-serpins remain to be further investigated using RNA interference and other molecular techniques.
Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.
This research is to determine the level of environmental pollution at a blasting construction area which is the origin of noise, vibration, and suspended particle, and to compare the results with other domestic and international standard data. This experiment is also to find out the effects resulting from blasting construction and to propose a plan that can decrease environmental pollution. The blasting construction area is a factory site which is about one and half million square meter and sewage disposal plant is about ninety thousand square meter. Both were selected as the areas for the tests to be conducted in determination test. The test to determine the level of noise, vibration, and total suspended particle was conducted thirty times around the blasting construction area by comparing measurement results and numerical analysis. However, as the test was not conducted in the laboratory but in the actual blasting construction area, it was not possible to do the test with the same exact conditions each true. Therefore, the test was not ideal as conditions could change from test to test. For the most part, the level of noise was below the standard level of 70dB. Every vibration test was under the standard limitation. For example, a house, 200m away was tested for noise and vibration and the level was found to be under the 0.2 cm/sec which is the standard for specialty designed cultural sites., i.e very low level. Also a buried oil pipeline that was 30m away also marked under 2.0cm/sec which is the norm for an industrial area. However, if there were an oil pipeline under the house, the amount of charging gunpowder per hole should be decreased compared to the amount used in the test. The test result for suspended particles under the standard limitation which is 24hour average 300$\mu\textrm{g}$/㎥ at a distance from blasting wavelength, but at detonator, total suspended particle from the blast origin exceeded the standard limitation. If explosion occurs when it detonates in the hole, most of the energy would be absorbed in the crushing of rocks, but some remaining energy would make noises and vibration inevitable. So the important thing is how to minimize the environmental pollution from the blasting. There should be regulations in order that the standard limitation is not exceeded, and to decrease the environmental pollution from the blasting.
Journal of Dental Rehabilitation and Applied Science
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v.27
no.4
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pp.393-403
/
2011
The purpose of this study was to analyze and compare survival rates of resorbable blast media(RBM) surface and acid-etched surface implants being usually used in clinics. RBM surface implants (USII, Osstem, Busan, Korea) or acid-etched surface implants ($Osseotite^{(R)}$, Biomet $3i^{[TM]}$, FL, USA) were placed in edentulous area of 140 patients between January of 2005 and March of 2007. The number of implants was 304, and 152 out of them were RBM surface implants while another 152 were acid-etched surface implants. According to the evaluation items, the survey was performed before and after the implants installations. The 3-year survival rates of both kind of implants were calculated. 1. Total of 152 RBM surface implants were placed. Among them, one implant was failed, which was implanted in the posterior mandible with D2 bone quality. The failure was resulted from fracture of the fixture. Others showed good results and survival rate of RBM surface implant was 99.34%. 2. Total of 152 acid-etched surface implants were placed. Seven implants of them were failed, thus, survival rate was 95.39%. The causes of the failures were considered as infection, overheat and the lack of initial stability. In this research, both implants showed good 3-year survival rate, although RMB surface implant represented a better result.
Rather than causing damage from heat, blast, and radiation of a regular nuclear weapon, recently, it is predicted that North Korea has been inventing high altitude electromagnetic pulse (HEMP) missile in order to incapacitate electronic equipment. HEMP shielding facility is used for military purpose today. Despite the electromagnetic shielding effects from high quality compression plates, problems may include such as the possibility of electromagnetic influx resulting in the welding of the compression plates, and difficulties and high cost of construction. Therefore, in this study, a high electrical conducting material was added to the concrete experimental subject to ensure the shielding effect through electromagnetic waves to for the concrete structure, instead of building a shielding facility separately for the structure. Also, among the experimental subjects, 100 ${\mu}m$ of Iron-Aluminum alloy metal spraying coat was applied to two types with the highest shielding effect, and to two types with the lowest shielding effect. The result of the experiment indicates that experimental subjects added with a high electrical conductivity material did not meet the minimum shielding criteria of MIL-STD-118-125-1 standard, but all the experimental material applied to the metal spraying coating satisfied the minimum shielding criteria. In conclusion, it is considered that 100 µm of Iron-Aluminum alloy metal spraying coat contains high efficiency in the HEMP shielding.
In the continued research on natural fungicides for control of plant diseases by using plant-derived products, we found that Spatholobus suberectus Dunn had a strong fungicidal activity against several plant pathogens. S. suberectus (1 kg) was extracted with 80% aqueous MeOH and then the concentrated extract was partitioned with n-hexane, EtOAc, n-BuOH and $H_2O$ successively. The four layers were tested their disease contron efficacies against 6 plant diseases such as rice blast (RCB), rice sheath blight (RSB), tomato grey mold (TGM), tomato late blight (TLB), wheat leaf rust (WLR), and barley powdery mildew (BPM). The EtOAc fraction was highly active showing over 80% control against RCB, TGM, TLB, and BPM. By using silica gel chromatography, preparative TLC and HPLC, six compounds that were expected to have antifungal activity were separated. Their chemical structures were identified as ethanone, hydroxytyrosol, epicatechin, procyanidin B2, dimethoxy daizein and formononetin by ESI-MS, $^1H$-NMR, $^{13}C$-NMR, and 2D-NMR spectroscopic analyses. The chemicals except epicatechin were first reported in S. suberectus. Study on in vitro and in vivo antifungal activities of the isolated compounds is in progress.
Full-length cDNAs are essential for the correct annotation of genomic sequences and for the functional analysis of genes and their products. To elucidate the functions of a large population of Chinese cabbage (Brassica rapa) genes and to search efficiently for agriculturally useful genes, we have been taking advantage of the full-length cDNA Over-eXpresser (FOX) gene hunting system. With oligo dT column it purify the each mRNA from the flower organs, leaf and stem tissue. And about 120,000 cDNAs from the library were transformed into $\lambda$-pFLCIII-F vector. Of which 115,000 cDNAs from the library were transformed into T-DNA binary vector, pBigs for transformation study. We used normalized full-length cDNA and introduced each cDNA into Arabidopsis by in planta transformation. Full-length Chinese cabbage cDNAs were expressed independently under the CaMV 35S promoter in Arabidopsis. Selfed seeds were harvested from transgenic Arabidopsis. We had selected 2,500 transgenic plants by hygromycin antibiotic tolerant test, and obtained a number of transgenic mutants. Each transgenic Arabidopsis was investigated in morphological changes, fertility and leaf colour. As a result, 285 possible morphological mutants were identified. Introduced cDNA was isolated by PCR amplification of the genomic DNA from the transgenic mutants. Sequencing result and BLAST analysis showed that most of the introduced cDNA were complete cDNAs and functional genes. Also, we examined the effect of Bromelain on enhancing resistance to soft rot in transgenic Chinese cabbage 'Osome'. The bromelain gene identified from FOX hunting system was transformed into Chinese cabbage using Agrobacterium methods. Transformants were screened by PCR, then RT-PCR and real time PCR were performed to analyze gene expression of cysteine protease in the T1 and T2 generations. The anti-bacterial activity of bromelain was tested in Chinese cabbages infected with soft rot bacteria. The results showed that the over-expressed bromelain gene from pineapple conferred enhanced resistance to soft rot in Chinese cabbage.
Climate in the year of 2001 could be summarized as severe drought from March to May, unusually high temperature in whole plant growth periods and clear weather condition especially in harvesting time of September and October without any typoons. In rice, major diseases such as leaf and panicle blast, bacterial blight, sheath blight and bacterial grain rot developed little due to unfavorable weather conditions, however, brown spot occurred severely due partly to the reduced use of N-fertilizer, Rice stripe virus infection was unusually high mainly in the west coast areas with four times more diseaseD area campared to the previous year, In pepper phytophthora blight was extremely severe in Cheonnam and Cheonbuk provinces, where had frequent rainfalls during growing period. Incidence of major diseases of tomato and cucumber in 2001 was relatively mild. In watermelon, penicillium fruit rot, one of the peculiar disease, spread over major production areas, whereas CGMMV, usually severe in every years was much reduced. Watermelon plants growing in open-fields were more severely diseased than those in plastic houses. Major diseases of chinese melon and strawberry occurred slightly and in particular, sudden wilt syndrome of chinese melon which was severe in 2000 showed mild infections in 2001. Incidence of white rot of garlic and onion was much lower unlike other years due mainly to spring drought and high temperature in the growing period. Infected area of potato bacterial wilt tended to increase in 2001 while fusarium wilt of sweetpotato was decreased. Apple diseases were generally mild, but powdery mildew of pear increased sharply. Barley scab was not severe as seen in other years.
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