• Title/Summary/Keyword: Genetic Level

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Transduced PEP-1-Grb7 Fusion Protein Suppressed LPS-induced COX-2 Expression

  • An, Jae-Jin;Kim, So-Young;Lee, Sun-Hwa;Kim, Dae-Won;Ryu, Hea-Jin;Yeo, Seung-Il;Jang, Sang-Ho;Kwon, Hyung-Joo;Kim, Tae-Yoon;Lee, Sang-Chul;Poo, Ha-Ryoung;Cho, Sung-Woo;Lee, Kil-Soo;Park, Jin-Seu;Eum, Won-Sik;Choi, Soo-Young
    • BMB Reports
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    • v.40 no.2
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    • pp.189-195
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    • 2007
  • Although the incidence and severity of atopic dermatitis (AD) is steadily increasing at an alarming rate, its pathogenic mechanisms remain poorly understood yet. Recently, we found that the expression of Grb7 protein was markedly decreased in AD patients using proteomic analysis. In the present study, human Grb7 gene was fused with PEP-1 peptide in a bacterial expression vector to produce a genetic in-frame PEP-1-Grb7 fusion protein. The expressed and purified PEP-1-Grb7 fusion proteins transduced efficiently into skin cells in a time- and dose-dependent manner when added exogenously in culture media. Once inside the cells, the transduced PEP-1-Grb7 protein was stable for 48 h. In addition, transduced PEP-1-Grb7 fusion protein markedly increased cell viability in macrophage RAW 264.7 cells treated with LPS by inhibition of the COX-2 expression level. These results suggest that the PEP-1-Grb7 fusion protein can be used in protein therapy for inflammatory skin disorders, including AD.

Identification and Phylogenetic Relationships of Inonotus obliquus Strains by PCR-RFLP of ITS sequences and STS markers (ITS 부위의PCR-RFLP 및 STS 마커를 이용한 차가버섯의 종 및 계통간 유연관계 분석)

  • Shin, Pyung-Gyun;Kong, Won-Sik;Yoo, Young-Bok;Lee, Keum-Hee;Oh, Se-Jong;Choi, Man-Soo
    • Journal of Mushroom
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    • v.7 no.4
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    • pp.150-155
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    • 2009
  • Because the import of Inonotus obliquus have been rapidly increased in Korea, we developed the Inonotus species-specific marker by using various sequences including ITS sequences, PCR-RFLP and STS primers and used this marker to determine both genetic relatedness and strains discrimination of Inonotus spp. Total 17 different Inonotus spp. were examined by using ITS sequences and classified into 2 different groups. One strain, ASI74008 isolated from Kamchaka island of Siberia, showed the high sequence identity (98%) at the nucleotide level to the other I. obliquus DSM strain, indicating the ASI74008 belong to I. obliquus species. Comparison of banding patterns after restriction enzyme digestions with PCR amplicons of ITS region revealed some variations depending on the species and strains. However, PCR products amplified with STS primer showed species specific patterns.Therefore, use of both STS primers and PCR-RFLP could help for better strain identification.

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Analysis of Single Nucleotide Polymorphism of eNOS Genes in Korean Genome (한국인의 eNOS 유전자 SNP 분석)

  • Lee, Hyung-Ran;Kim, Su-Won;Yoo, Min
    • Journal of Life Science
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    • v.24 no.2
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    • pp.181-185
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    • 2014
  • We identified SNPs (single nucleotide polymorphisms) for endothelial nitric oxide synthase (eNOS) genes in the Korean genome. eNOS is present in the vascular endothelium, platelets, and several other cell types that continuously produce modest amounts of NO. Endothelium-derived NO plays a key role in the regulation of vascular tone, and the impaired effects of NO on the cardiovascular system appear to be responsible for coronary atherosclerosis and thrombosis. In recent studies, a missense variant within exon 7 of the eNOS gene in patients with coronary spastic angina-GAG to GAT substitution, which results in the replacement of glutamic acid by aspartic acid (Glu298Asp [G894T])-has been identified and is known to be significantly associated with coronary spasm. We prepared PCR primers based on sequences in Genbank. Primers were prepared for normal and SNPs separately, as reported for other Asian countries, such as G894T. Their sequences were different only at the 3' ends so that primer extension could only by possible when base pairs between templates and primers matched. We also employed ARMS (Amplification Refractory Mutation System) technology to improve the specificity of the PCR reaction. In conclusion, we were able to demonstrate the eNOS G894A polymorphism in Korean gemone. This study should facilitate research on the cause of myocardial infarction and development on further therapy at the genetic level.

Distribution and Toxin Gene Characteristic of Bacillus cereus Isolated from Foods in Busan (부산지역 가공식품 중 Bacillus cereus 분포 현황 및 독소 유전자 특성)

  • Park, So-hyun;Gwon, Wi-Gyeong;Lee, In-sook;Kim, Eun-ju;Hwang, Su-jeong;Koo, Hee-soo;Na, Young-ran;Kim, Byung-jun;Park, Eun-hee;Lee, Mee-ok
    • Journal of Food Hygiene and Safety
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    • v.35 no.3
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    • pp.219-224
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    • 2020
  • The purpose of this study was to provide basic data necessary for the prevention of food poisoning and safe food management. We examined 872 food samples for B. cereus in accordance with the MFDS Food Code and investigated characteristics of their harboring toxin genes. We detected and isolated 113 strains of B. cereus from 78 food samples (8.9%), and the average detection level was 48 CFU/g. B. cereus isolates carried at least 1 toxin gene among the emetic toxins and 5 enterotoxin genes. The toxin gene profiles of B. cereus were classified into 18 different types of isolates showing genetic diversity. Among the strains, 34 (30.1%) had all 5 enterotoxin genes (Cytk-nheA-entFM-bceT-hblC), accounting for the highest percentage. The entFM and nheA genes were major enterotoxin genes, while the emetic toxin gene, CER, was the least detected in B. cereus isolated from food samples.

Genomic Diversity of Helicobacter pylori

  • Lee, Woo-Kon;Choi, Sang-Haeng;Park, Seong-Gyu;Choi, Yeo-Jeong;Choe, Mi-Young;Park, Jeong-Won;Jung, Sun-Ae;Byun, Eun-Young;Song, Jae-Young;Jung, Tae-Sung;Lee, Byung-Sang;Baik, Seung-Chul;Cho, Myung-Je
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.519-532
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    • 1999
  • Helicobacter pylori is a causative agent of type B gastritis and plays a central role in the pathogenesis of gastroduodenal ulcer and gastric cancer. To elucidate the host-parasite relationship of the H. pylori infection on the basis of molecular biology, we tried to evaluate the genomic diversity of H. pylori. An ordered overlapping bacterial artificial chromosome (BAC) library of a Korean isolate, H. pylori 51 was constructed to set up a genomic map. A circular physical map was constructed by aligning ApaI, NotI and SfiI-digested chromosomal DNA. When the physical map of H. pylori 51 was compared to that of unrelated strain, H. pylori 26695, completely different restriction patterns were shown. Fifteen known genes were mapped on the chromosome of H. pylori 51 and the genetic map was compared with those of strain 26695 and J99, of which the entire genomic sequences were reported. There were some variability in the gene location as well as gene order among three strains. For further analysis on the genomic diversity of H. pylori, when comparing the genomic structure of 150 H. pylori Korean isolates with one another, genomic macrodiversity of H. pylori was characterized by several features: whether or not susceptible to restriction digestion of the chromsome, variation in chromosomal restriction fingerprint and/or high frequency of gene rearrangement. We also examined the extent of allelic variation in nucleotide or deduced amino acid sequences at the individual gene level. fucT, cagA and vacA were confirmed to carry regions of high variation in nucleotide sequence among strains. The plasticity zone and strain-specific genes of H. pylori 51 were analyzed and compared with the former two genomic sequences. It should be noted that the H. pylori 51-specific sequences were dispersed on the chromosome, not congregated in the plasticity zone unlike J99- or 26695-specific genes, suggesting the high frequency of gene rearrangement in H. pylori genome. The genome of H. pylori 51 shows differences in the overall genomic organization, gene order, and even in the nucleotide sequences among the H. pylori strains, which are far greater than the differences reported on the genomic diversity of H. pylori.

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Occurrence of Fusarium Species in Korean Sorghum Grains (국내 수수 알곡에서의 Fusarium속 균의 발생현황)

  • Choi, Jung-Hye;Nah, Ju-Young;Jin, Hyun-Suk;Lim, Su-Bin;Paek, Ji-Seon;Lee, Mi-Jeong;Jang, Ja-Yeong;Lee, Theresa;Hong, Sung Kee;Choi, Hyo-Won;Kim, Jeomsoon
    • Research in Plant Disease
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    • v.25 no.4
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    • pp.213-219
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    • 2019
  • A total of 1,159 Fusarium strains were isolated from sorghum grown in Danyang and Youngwol in 2017 and 2018. The isolates were analyzed to reveal genetic, toxigenic and pathogenic characteristics. Phylogenetic analysis using TEF-1α and RPB2 genes showed that the samples were contaminated with at least 17 Fusarium species. Among them, F. graminearum, F. proliferatum, F. thapsinum, F. incarnatum, and F. asiaticum were dominant species. In F. graminearum and F. asiaticum, F. graminearum-15-acetyl deoxynivalenol chemotype and F. asiaticum-nivalenol chemotype were frequent. Six Fusarium species tested produced one or more mycotoxins, except F. thapsinum and FTSC 11. F. proliferatum and F. fujikuroi had FUM1 gene (76.0% and 81.6%, respectively) and some isolates produced high level of fumonisin (over 1,000 ㎍). F. proliferatum and F. thapsinum were more virulent than other species on sorghum. These results indicate that Fusarium species in sorghum might produce multiple mycotoxins.

Characterization of the drug receptors responsible for intestinal contraction in Israeli carp (이스라엘잉어 장관 수축에 관여하는 약물수용체에 관한 연구)

  • Yun, Hyo-in;Han, Kyong-oh;Park, Seung-chun;Cho, Joon-hyoung;Oh, Tae-kwang
    • Korean Journal of Veterinary Research
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    • v.34 no.3
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    • pp.447-456
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    • 1994
  • In order to elucidate the characterization of receptors involved in inestinal motility of Israeli carp, spontaneously contracting Israeli carp intestinal preperations were prepared and mounted in the organ chambers for contraction traicings using a polygraph. Various contractile agonists were treated and their dose-response curves were constructed. $EC_{50}$ values$(pD_2)$ of each agonist on specific receptors, $pA_2$ values of competitive antagonists against some agonists, and $K_1$, values of noncompetitive antagonists against some agonists were analyzed for characterization of receptors related with the intestinal contraction. Results obtained through the experiments were summarized as follows: 1. Acetylcholine(ACh) exhibited biphasic dose-response curves: initial ACh-induced dose dependent contractions were observed in pM levels but followed by decreased response in in-between concentration levels. Dose dependent contractions reappeared in ${\mu}M$ level. The peaks in pM and ${\mu}M$ levels appeared in $10^{-13}M$ and $3{\times}10^{-5}M$, respectvely. 2. Carbachol(CaCh) exhibited dose dependent contractions from $10^{-9}M$ to $10^{-5}M$, and its $pD_2$ values were higher than those of ACh($5.60{\pm}0.11$). ACh and CaCh exhibited equiactive contractions. Nicotine had no effects on contractile responses of Israeli carp intestine. 3. ACh-induced responses were inhibited by atropine($K_1:7{\times}10^{-8}M$), a muscarinic antagonist, in a non-competitive manner. But CaCh-induced responses were inhibited by both antimuscarinic atropine($pA_2:9.52{\pm}0.14$) and selective $M_2$ antagonistic 4-DAMP($pA_2:8.16{\pm}0.09$), in competitive manners. Nicotine receptor antagonistic decamethonium and hexamethonium had no effects on ACh-and CaCh-induced contractions. Therefore, the cholinergic receptor related to intestinal motility of Israeli carp was assumed as $M_2$ type. 4. In Israeli carp intestine, 5-HT (serotonin) exhibited dose dependent contractions in concentration range from $10^{-8}M$ to $10^{-5}M$. The maximal responses, however, were corresponded to about 50% of those of ACh or CaCh. 5-HT induced contractions were inhibited by $5-HT_2$ antagonistic ketanserin ($K_1: 7.8{\times}10^{-4}M$) in a non-competitive manner, but not by both of anti $5-HT_1$, spiperone and anti $5-HT_3$, MDL-72222. Hence, $5-HT_2$ receptors are suggested to be existed in Isreli carp intestine.

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Functional Expression of Anti-BNP scFv in E. coli Cytoplasm for the Detection of B-type Natriuretic Peptide (B-type natriuretic peptide 분석을 위한 항 BNP scFv 항체의 대장균 세포질 내에서의 기능적 발현)

  • Maeng, Bo-Hee;Nam, Dong-Hyun;Kim, Yong-Hwan
    • KSBB Journal
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    • v.24 no.6
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    • pp.591-597
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    • 2009
  • B-type natriuretic peptide is a neurohormone secreted in the cardiac ventricles. BNP levels are elevated in patients with ventricular dysfunction. Therefore, the concentration of BNP is important factor to reflect diagnosis and prognosis for cardiovascular disease. In this respect, anti-BNP scFv is an urgent requirement for early diagnosis in the field of biosensor. Herein, the genetic codes of anti-BNP scFv were chemically synthesized and cloned into both pET22b (+) and pColdⅣ vector, respectively. The recombinant scFv was successfully expressed as a functional form in cytoplasm of E. coli and detected through Western blot and ELISA. The highest level of functional expression of anti-BNP scFv was achieved using pET22b (+) vector at $15^{\circ}C$ by addition of 0.1 mM IPTG. Additionally, being exposed to both BNP and ANP, anti-BNP scFv specifically captured only BNP. Therefore, anti-BNP scFv expressed in this study will be applied to measure the concentration of BNP as a diagnostic recognition molecule.

Inheritance and Heritability of Telomere Length in Chicken (닭 텔로미어 길이의 유전력 추정과 유전 전이 양상)

  • Park, Dan Bi;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
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    • v.41 no.3
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    • pp.217-225
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    • 2014
  • Telomeres are the ends of the eukaryotic chromosomes and consist of a tandem repetitive DNA sequence and shelterin protein complex. The function of telomere is to protect chromosome. Telomere length in somatic cells tends to decrease with organismal age due to the end replication problem. However, several factors at the genetic, epigenetic and environmental level affect telomere length. In this study, we estimated heritability of telomere length and investigated inheritance of telomeres in a chicken. Telomere length of lymphocytes was analyzed by semi-quantitative polymerase chain reaction using telomere primer and quantitative fluorescence in situ hybridization using telomeric DNA probe. In results, heritability of telomere length was estimated 0.9 at birth by offspring-parent regression analysis and was estimated 0.03 and 0.04 at 10 and 30 weeks old, respectively, by parental variance analysis. There was a significant positive correlation in telomere length between father and their offspring (r=0.348), and mother and their offspring (r=0.380). In inheritance patterns of telomere length, the influence of paternal and maternal effect on their offspring was similar. The influence of inherited telomeres on male and female progeny was also roughly alike. These results implicated that imprinting of parental telomere length was regulated by autosomal genes, not sex linked genes. In addition, telomere length of offspring at birth did not differ along with their maternal age. Thus, maternal age does not affects telomere length in their offspring at birth owing to cellular reprogramming at early embryonic stage.

Study on Effects of Population Stratification on Haplotype Trend Test in Case-Control Studies (환자-대조군 연구에서 인구집단 층화가 일배체형 경향성 검정에 미치는 영향)

  • Kim, Jin-Heum;Kang, Dae-Ryong;Lim, Hyun-Sun;Nam, Chung-Mo
    • The Korean Journal of Applied Statistics
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    • v.22 no.5
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    • pp.1085-1096
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    • 2009
  • Population stratification can cause spurious associations between genetic markers and disease locus. In order to handle this population stratification in haplotype-based case-control association studies, we added population indicators as covariates to the haplotype trend regression model proposed by Zaykin et al. (2002). We investigated through simulations how both population stratification and measurement error in the estimation of true population of each individual affect type I error probabilities of the association tests based on both Zaykin et al.'s (2002) model and the proposed model. Based on those results, in the situation that there exists population stratification but there is no error in population classification of each individual, our proposed model does satisfy a type I error probability whereas Zaykin et al.'s (2002) model does not. However, as the measurement error increases, a type I error probability of our model correspondingly becomes larger than a nominal significance level. It implies that as long as uncertainty in the estimation of true population of each individual still remains, it is nearly impossible to avoid false positive in case-control association studies based on haplotypes.