• Title/Summary/Keyword: specific genes

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Analysis of Molecular Epidemiological Properties of Staphylococcus aureus Isolates from Domestic Animals and Human Patients by PCR (Polymerase Chain Reaction을 활용한 국내 동물과 사람환자에서 분리한 Staphylococcus aureus 분리주의 분자역학적 특성분석)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.24-37
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    • 2005
  • This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains $(100\%)$ tested and followed by coa-gene $(87.9\%)$, spa-gene $(91.4\%)$ and aroA-gene $(26.7\%)$, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains $(12.1\%)$ were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.

Aberrant Promoter Methylation of Death-Associated Protein Kinase in Serum DNA from Lung Cancer Patients (원발성 폐암 환자의 혈청에서 DAP kinase 유전자의 Methylation 양상)

  • Lee, Jun Hee;Lee, Jung Wook;Jung, Kyung Sik;Kim, Ki Uk;Lee, Tae Kun;Lee, Kyung Woo;Na, Min-Ah;Jeon, Doo Soo;Choi, Young Min;Kim, Yun Seong;Lee, Min Ki;Park, Soon Kew
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.4
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    • pp.378-387
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    • 2003
  • Background : Promoter methylation of tumor suppressor genes is one of the key epigenetic changes in many human cancers. The aim of this study was to evaluate the promoter methylation status of the Death-associated protein(DAP) kinase gene, which played an important role in lung cancer, in the serum DNA of primary lung cancer patients. Methods : This study investigated the aberrant methylation of DAP kinase in the serum of 65 primary lung cancer patients by methylation-specific PCR (MSP). Results : Methylation in the serum was detected in 29 of 65(44.6%) for DAP kinase. There was no statistical association between methylation of DAP kinase and age, smoking history, histologic type, or stage. Methylation of DAP kinase was found more frequently in men (p=0.044). Conclusions : This study suggests that the aberrant methylation of the DAP kinase promoter is readily detectable in the serum DNA of lung cancer patients using MSP analysis.

Characterization of Traits Related to Grain Shape in Korean Rice Varieties (국내 육성 벼 품종 입형 관련 특성 분석)

  • Lee, Chang-Min;Lee, Keon-Mi;Baek, Man-Kee;Kim, Woo-Jae;Suh, Jung-Pil;Jeong, Oh-Young;Cho, Young-Chan;Park, Hyun-Su;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.199-213
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    • 2020
  • Grain size and shape are the two important components contributing to rice yield and quality. To analyze traits related to grain-shape, a total of 272 varieties derived from japonica, japonica black and Tongil-type rice accession in Korea were evaluated in this study. The traits, grain length (GL), grain width (GW), grain thickness (GT), length to width ratio (RLW), and 1000-grain weight (TGW) were measured and replicated 10 times. Genes (GW2, GS3, qGL3, qSW5, GS5, TGW6, GW7, and GW8) related to grain-shape were validated in the accessions using specific DNA marker sets. K-mean clustering of the accession based on phenotypic data revealed three groups: group 1 was classified by GW and GT and included most of japonica type, group 2 was classified by RLW and GL reached a medium size and possessed a half spindle-shaped type, and group 3 was classified by TGW, reached a long size and possessed a semi-round shape. In validation tests using the marker sets, both gw2 and tgw6 were validated in less than 1% of the tested accessions and two allelic types, qgl3 and gw8, were only verified in Tongil-type accessions. For GW8 and GW2, any different amplicons were not amplified in any japonica or Tongil-type accessions, respectively. In order to suggest the representative grain-shape gene combinations for each ecotype, the allelic combinations were evaluated by PCR analysis. Cj1 and 2 in japonica (Cj1-7), Cj_b1 and 2 in japonica-black (Cj_b1-3), and CT3 in Tongil-type (CT1-13) turned out to be the dominant combination in each ecotype, respectively. In addition, the results revealed that introgression of four genes (gw2, gs3, qSW5, and GS5) would expand the diversity of grain shape in Korean japonica varieties. The gene combinations information could be utilized practically to understand or enhance grain shape in japonica rice breeding program.

Studies on Ecological Variation and Inheritance for Agronomical Characters of Sweet Sorghum Varieties (Sorghum vulgare PERS) in Korea (단수수(Sorghum vulgare PERS) 품종의 생태변이 및 유용형질의 유전에 관한 연구)

  • Se-Ho Son
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.10
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    • pp.1-43
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    • 1971
  • Experiment I: The objective of this study was to know variation in some selected agronomic characters of sweet sorghum when planted in several growing seasons. The 17 different sweet sorghum varieties having various maturities, and plant, syrup and sugar types were used in this study which had been carried out for the period of two years from 1968 to 1969 at Industrial Crops Division of Crop Experiment Station in Suwon. These varieties were planted at an interval of 20 days from April 5 to August 25 both in 1968 and 1969. The experimental results could be summarized as follows: 1. As planting was made early, the number of days from sowing to germination was getting prolonged while germination took place early when planted at the later date of which air temperature was relatively higher. However, such a tendency was not observed beyond the planting on August 25. In general, a significant negative correlation was found between the number of days from sowing to germination and the average daily temperature but a positive correlation was found between the former and the total accumulated average temperature during the growth period. 2. The period from sowing to heading was generally shortened as planting was getting delayed. The average varietal difference in number of days from sowing to heading was as much as 30.2 days. All the varieties were grouped into early-, medium and late-maturing groups based upon a difference of 10 days in heading. The average number of days from sowing to heading was 78.5$\pm$4.5 days in the early-maturing varieties, 88.5$\pm$4.5 days in the medium varieties and 98.5$\pm$4.5 days in the late-maturing varieties, respectively. The early-maturing varieties had the shortest period to heading when planted from July 15 to August 5, the medium varieties did when planted before July 15 and the late-maturing varieties did when planted before June 5. 3. The relationship between the sowing date (x) and number of days from sowing to heading could be expressed in an equation of y=a+bx. A highly positive correlation was found between the coefficient of the equation(shortening rate in heading time) and the average number of days from sowing to heading. 4. The number of days from sowing to heading was shortened as the daily average temperature during the growth period was getting higher. Early-maturing varieties had the shortest period to heading at a temperature of 24.2$^{\circ}C$, medium varieties at 23.8$^{\circ}C$ and late-maturing varieties at 22.9$^{\circ}C$, respectively. In other words, the number of days from sowing to heading was shortened rapidly in case that the average temperature for 30 days before heading was 22$^{\circ}C$ to $25^{\circ}C$. It prolonged relatively when the temperature was lower than 21$^{\circ}C$. 5. There was a little difference in plant height among varieties. In case of early planting, no noticeable difference in the height was observed. The plant height shortened generally as planting season was delayed. Elongation of plant height was remarkably accelerated as planting was delayed. This tendency was more pronounced in case of early-maturing varieties rather than late-maturing varieties. As a result, the difference in plant height between the maximum and the minimum was greater in late-maturing varieties than in early-maturing varieties. 6. Diameter of the stalk was getting thicker as planted earlier in late-maturing varieties. On the other hand, medium or early-maturing varieties had he thickest diameter when they were planted on April 25. 7. In general, a higher stalk yield was obtained when planted from April 25 to May 15. However, the planting time for the maximum stalk yield varied from one variety to another depending upon maturity of variety. Ear]y-maturing varieties produced the maximum yield when planted about April 25, medium varieties from April 25 to May 15 and late-maturing varieties did when planted from April 5 to May 15 respectively. The yield decreased linearly when they were planted later than the above dates. 8. A varietal difference in Brix % was also observed. The Brix % decreased linearly when the varieties were planted later than May 15. Therefore, a highly negative relationship between planting date(x) and Brix %(y) was detected. 9. The Brix % during 40 to 45 days after leading was the highest at the 1st to the 3rd internodes from the top while it decreased gradually from the 4th internode. It increased again somewhat at the 2nd internode from the ground level. However, it showed a reverse relationship between the Brix % and position of internode before heading. 10. Sugar content in stalk decreased gradually as planting was getting delayed though one variety differed from another. It seemed that sweet sorghum which planted later than June had no value as a sugar crop at all. 11. The Brix % and sugar content in stalk increased from heading and reached the maximum 40 to 45 days after heading. The percentage of purity showed the same tendency as the mentioned characters. Accordingly, a highly positive correlation was observed between. percentage of purity and Brix % or sugar content in stalk. 12. The highest refinable sugar yield was obtained from the planting on April 25 in late-maturing varieties and from that on May 15 in early-maturing varieties. The yield rapidly decreased when planted later than those dates. Such a negative correlation between planting date(x) and refinable sugar yield(y) was highly significant at 1% level. 13. Negative correlations or linear regressions between delayed planting and the number of days from sowing to germination. accumulated temperature during germination period, number of days to heading, accumulated temperature to heading, plant height, stem diameter, stalk weight, Brix %. sugar content, refinable sugar yield or Purity % were obtained. On the other hand, highly positive correlations between the number of days from sowing to heading(x) and Brix %, sugar content, purity %, refinable sugar yield, plant height or stalk yield, between Brix %(x) and purity %, refinable sugar yield or stalk yield, between sugar content(x) and purity% or refinable sugar yield(y), between purity %(x) and refinable sugar yield and between daylength at heading(x) and Brix %. number of days from sowing to heading, sugar content, purity % or refinable sugar yield (y), were found, respectively. Experiment II: The 11 varieties were selected out of the varieties used in Experiment I from ecological and genetic viewpoints. Complete diallel cross were made among them and the heading date, stalk length, stalk yield, Brix %, syrup yield, combining ability and genetic behavior of F$_1$ plants and their parental varieties were investigated. The results could be summarized as follows: 1. In general, number of days to heading showed a partial dominance over earliness or late maturity or had a mid-value, though there were some specific combinations showing a complete dominance or transgressive segregation in maturity. Some combinations showed relatively high general or specific combining abilities in maturity. Therefore, a 50 to 50 segregation ratio in heading date could be estimated in this study and it might be positive to have a selection in early generation since heritability of the character was relatively high. 2. A vigorous hybrid vigor was observed in stalk length. A complete or partial dominant effect of long stalk was obtained. The general combining ability and specific combining ability of stalk length were generally high. Long and short stalks segregated in a ratio of 50:50 and its heritability was relatively low. 3. Except for several specific combinations, high stalk yield seemed to be partial dominant over the low yield. Some varieties demonstrated relatively high general as well as specific combining abilities. It was assumed that several recessive genes were involved in expression of this character. The interaction among regulating recessive genes was also obtained. Accordingly, the heritability of stalk yield seemed to be rather low. 4. The Brix % of hybrid plants located around mid-parental value though some of them showed much higher or lower percentage. It could be explained by the fact that such behavior might be due to partial dominance of Brix %. The varieties with, relatively higher Brix % were high both in general. and specific combining abilities. Therefore, it could be recommended to use the varieties having higher sugar content in order to develop higher-sugar varieties. 5. The syrup yield seemed to be transgressively segregated or completely dominant over low yield. Hybrid vigor of syrup yield was relatively high. No-consistent relationship between general combining ability and specific combining ability was observed. However, some cases demonstrated that the varieties with relatively higher general combining ability had relatively lower specific combining ability. It was assumed that the frequencies of dominant and recessive alleles were almost same.

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Effects of climate change on biodiversity and measures for them (생물다양성에 대한 기후변화의 영향과 그 대책)

  • An, Ji Hong;Lim, Chi Hong;Jung, Song Hie;Kim, A Reum;Lee, Chang Seok
    • Journal of Wetlands Research
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    • v.18 no.4
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    • pp.474-480
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    • 2016
  • In this study, formation background of biodiversity and its changes in the process of geologic history, and effects of climate change on biodiversity and human were discussed and the alternatives to reduce the effects of climate change were suggested. Biodiversity is 'the variety of life' and refers collectively to variation at all levels of biological organization. That is, biodiversity encompasses the genes, species and ecosystems and their interactions. It provides the basis for ecosystems and the services on which all people fundamentally depend. Nevertheless, today, biodiversity is increasingly threatened, usually as the result of human activity. Diverse organisms on earth, which are estimated as 10 to 30 million species, are the result of adaptation and evolution to various environments through long history of four billion years since the birth of life. Countlessly many organisms composing biodiversity have specific characteristics, respectively and are interrelated with each other through diverse relationship. Environment of the earth, on which we live, has also created for long years through extensive relationship and interaction of those organisms. We mankind also live through interrelationship with the other organisms as an organism. The man cannot lives without the other organisms around him. Even though so, human beings accelerate mean extinction rate about 1,000 times compared with that of the past for recent several years. We have to conserve biodiversity for plentiful life of our future generation and are responsible for sustainable use of biodiversity. Korea has achieved faster economic growth than any other countries in the world. On the other hand, Korea had hold originally rich biodiversity as it is not only a peninsula country stretched lengthily from north to south but also three sides are surrounded by sea. But they disappeared increasingly in the process of fast economic growth. Korean people have created specific Korean culture by coexistence with nature through a long history of agriculture, forestry, and fishery. But in recent years, the relationship between Korean and nature became far in the processes of introduction of western culture and development of science and technology and specific natural feature born from harmonious combination between nature and culture disappears more and more. Population of Korea is expected to be reduced as contrasted with world population growing continuously. At this time, we need to restore biodiversity damaged in the processes of rapid population growth and economic development in concert with recovery of natural ecosystem due to population decrease. There were grand extinction events of five times since the birth of life on the earth. Modern extinction is very rapid and human activity is major causal factor. In these respects, it is distinguished from the past one. Climate change is real. Biodiversity is very vulnerable to climate change. If organisms did not find a survival method such as 'adaptation through evolution', 'movement to the other place where they can exist', and so on in the changed environment, they would extinct. In this respect, if climate change is continued, biodiversity should be damaged greatly. Furthermore, climate change would also influence on human life and socio-economic environment through change of biodiversity. Therefore, we need to grasp the effects that climate change influences on biodiversity more actively and further to prepare the alternatives to reduce the damage. Change of phenology, change of distribution range including vegetation shift, disharmony of interaction among organisms, reduction of reproduction and growth rates due to odd food chain, degradation of coral reef, and so on are emerged as the effects of climate change on biodiversity. Expansion of infectious disease, reduction of food production, change of cultivation range of crops, change of fishing ground and time, and so on appear as the effects on human. To solve climate change problem, first of all, we need to mitigate climate change by reducing discharge of warming gases. But even though we now stop discharge of warming gases, climate change is expected to be continued for the time being. In this respect, preparing adaptive strategy of climate change can be more realistic. Continuous monitoring to observe the effects of climate change on biodiversity and establishment of monitoring system have to be preceded over all others. Insurance of diverse ecological spaces where biodiversity can establish, assisted migration, and establishment of horizontal network from south to north and vertical one from lowland to upland ecological networks could be recommended as the alternatives to aid adaptation of biodiversity to the changing climate.

BCR (Brown Color Repressor) gene isolation related to mycelial browning of Lentinus edodes (표고균사 갈변과 관련된 BCR (Brown Color Repressor) 유전자 분리)

  • Kim, Young-Ho;Park, Soo-Cheol;Jhune, Chang-Sung;You, Chang-Hyun;Sung, Jae-Mo;Kong, Won-Sik
    • Journal of Mushroom
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    • v.10 no.3
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    • pp.120-128
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    • 2012
  • Recently sawdust cultivation of Shiitake mushroom (Lentinula edodes) is increasing. It is important to make mycelia to be brown on the substrate surface. This browned surface in sawdust cultivation plays an important role like as artificial bark of the oak log, which protects the other pests and suppresses water evaporation in the substrate. In order to isolate genes which related to brown color formation, differential display method was used. Two cDNA fragments obtained by DD-PCR were 1.2 and 1.6kb and these were expressed in white colored mycelia from L. edodes, but not brown colored mycelia. Partial sequencing of these cDNA fragments showed that the 1.6kb cDNA had 100% identity with the microsatellites gene from Dugenia polichroa. However, the other 1.2kb cDNA fragment had poly T tail on 3' region of partial open reading frame on 5' region. The new primer designed based on the sequence of 1.2kb cDNA was constructed. RT-PCR analysis using the newly designed 0.12kb cDNA specific primer showed that the gene was only expressed in white color mycelia, but not in brown color mycelia. Sequence analysis of 5' region of this 1.2kb cDNA revealed that this gene contained partial open reading frame consisted of 110 amino acid. Homology search using DNASIS database showed that this gene had high sequence homology of 66.7% in DNA level and 69.2 % in amino acid level with dTDP-glucose 4,6-dehydratases gene from Arabidopsis thaliata. The dTDP-glucose 4,6-dehydratases gene was known to be function to have tolerance with oxidation stress. These results strongly suggest that this gene isolated from white mycelia of L. edodes might have a function of repressor against mycelia browning. Therefore I designated this gene as BCR (Brown Color Repressor) gene.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • 2000.10a
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. The subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Prevalence Report of Transovarian Transmitted Diseases in the Breeder Chickens, Korea (국내 종계에서 난계대 전염병 감염 실태 보고)

  • Kwon, Yong-Kuk;Kang, Min-Soo;Oh, Jae-Young;Jung, Byeong-Yeal;Kim, Hye-Ryoung;Kim, Ha-Young;Shin, So-Yeon;Kwon, Jun-Hun;Chung, Gab-Soo
    • Korean Journal of Poultry Science
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    • v.37 no.3
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    • pp.237-245
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    • 2010
  • A survey of transovarially transmitted diseases, including salmonellosis [(pullorum disease; PD)/(fowl typhoid; FT)], mycoplasmosis, avian infectious anemia (CIA), and fowl adenovirus infection was conducted in the breeder chickens from August to December in 2009. The numbers of flocks sampled out were: Grand Parents Stock (GPS), 45; Parents Stock (PS) 1,018, Baeksemi breeder (BSB) 54. The seroprevalence of salmonellosis (PD/FT) were 0% (GPS), 3.2% (PS), and 3% (BSB), respectively. A total of 983 chicken farms were affected with FT outbreaks between 2000 and 2008. The incidence of FT in commercial broilers, Baeksemi, commercial layers, native chickens, and broiler breeders was 44.3%, 26.2%, 15.7%, 12.6% and 1.08%, respectively. Of the affected broilers, over 90% birds were under 2 weeks of age, indicating it was possible that they were infected with S. gallinarum via vertical transmission. The sero-positive flocks against Mycoplasma gallisepticum (MG) were 71.1% (GPS), 88.7% (PS), 88.7% (BSB), while the rates of positive flocks against Mycoplasma synoviae (MS) were 86.0% (GPS), 77.0% (PS), and 98.0% (BSB). In GP and parent farms, the detection rates on specific genes of CIA virus were 19/45 (42.2%), and 169/1039 (18.0%), respectively, whereas the seroprevalence of CIA were 86.0% in GPS and 93.7% in PS flocks. In addition, positive flocks of fowl adenoviruses were 4.4% (GPS), 2.7% (PS) and 9.35% (BSB), respectively. As the results, avian mycoplasmosis and CIA have been more prevailing in chicken breeder than avian salmonellosis and fowl adenovirus infection in Korea.

Substrate chain-length specificities of polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas aeruginosa P-5 (Pseudomonas aeruginosa P-5에 존재하는 polyhydroxyalkanoate synthase PhaC1과 PhaC2의 기질특이성)

  • Woo, Sang Hee;Lee, Sun Hee;Rhee, Young Ha
    • Korean Journal of Microbiology
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    • v.52 no.4
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    • pp.455-462
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    • 2016
  • Pseudomonas aeruginosa P-5 is an unusual organism capable of synthesizing polyhydroxyalkanoates (PHAs) consisting of 3-hydroxyvalerate (3HV) and medium-chain-length (MCL) 3-hydroxyalkanoate (3HA) monomer units when C-odd alkanoic acids are fed as the sole carbon source. Evaluation of the substrate chain-length specificity of two P. aeruginosa P-5 PHA synthases ($PhaC1_{P-5}$ and $PhaC2_{P-5}$) by heterologous expression of $PhaC1_{P-5}$ and $PhaC2_{P-5}$ genes in Pseudomonas putida GPp104 revealed that $PhaC2_{P-5}$ incorporates both 3HV and MCL 3HAs into PHA, whereas $PhaC1_{P-5}$ favors only MCL 3HAs for polymerization. In order to obtain $PhaC2_{P-5}$ mutants with altered substrate specificity, site-specific mutagenesis for $PhaC2_{P-5}$ was conducted. Amino acid substitutions of $PhaC2_{P-5}$ at two positions (Ser326Thr and Gln482Lys) were very effective for synthesizing copolymers with a higher 3HV fraction. When recombinant P. putida GPp104 harboring double mutated $phaC2_{P-5}$ gene ($phaC2_{P-5}QKST$) was grown on nonanoic acid, 2.5-fold increase of copolymer content with 3.8-fold increase of 3HV fraction was observed. The $phaC2_{P-5}QKST$-containing Ralstonia eutropha PHB-4 supplemented with valeric acid also produced copolymers consisting of 3HV and 3-hydroxyheptanoate with a high 3HV fraction. These results suggest that recombinants containing $phaC2_{P-5}QKST$ could be useful for production of new PHA copolymers with improved material properties.

The Distribution of ${\gamma}{\delta}$ T Cells in Tuberculous Lymphadenopathy (결핵성 림프절에서 ${\gamma}{\delta}$ T 림프구의 분포에 관한 연구)

  • Shim, Tae-Sun;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.5
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    • pp.484-488
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    • 1994
  • Background : The antigen-specific receptor on the surface of most peripheral T lymphocytes is a disulfide-linked heterodimer composed of $\alpha$ and $\gamma$ subunits, noncovalently associated with CD3 polypeptides. Recently, a novel type of CD3-associated heterodimer was described on a T cell subset that does not express CD4 or CD8 molecules. This second type of TCR dimer is composed of chains encoded for by the $\gamma$- and $\delta$-TCR genes. These cells may exert both cytotoxic and lymphokine producing functions. Although it was reported that some ${\gamma}{\delta}$-TCR might recognize an MHC-linked determinant, the funεtion or physiologic ligand for this new receptor is not yet clear. It was found that ${\gamma}{\delta}$-TCR can react with 65 kD heat shock protein of M. tuberculosis, which suggests the possible protective role of ${\gamma}{\delta}$ T lymphocytes against tuberculosis. In our previous study, there was neither the increase in number nor the functional activation of ${\gamma}{\delta}$ T cells in the peripheral blood from patients with pulmonary tuberculosis. Now we report the distribution of ${\gamma}{\delta}$ T cells in the regional sites of M. tuberculosis infection, especial1y tuberculous lymphadenitis. Methods : Lymph nodes from patients with pathologically-proven tuberculous lymphadenopathy (n=5) and reactive hyperplasia (n=3) were used. Tissues were frozen in liquid nitrogen immediately after removal and stored below $-70^{\circ}C$. The cryostat sections of these frozen specimens were stained with anti-Leu-4 Ab, Identi-T TCR ${\delta}1$, and Identi-T ${\beta}F1$. The number of positively stained cells were counted at high power field. Results : The infiltration of ${\gamma}{\delta}$ T cells was significantly higher in the lymph nodes from patients with tuberculous lymphadenopathy than that with reactive hyperplasia ($16.3{\pm}10.3%$ vs. $1.7{\pm}1.5%$). Conclusion : These results suggest that ${\gamma}{\delta}$) T cells may play a role in the defense against M. tuberculosis infection, especially in the regional sites of infection.

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