• Title/Summary/Keyword: p63 gene

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Optimal Culture Conditions and Isolation of a ι-Carrageenase-producing Marine Bacterium

  • Shim Hang-Sun;Lim Su-Jin;Choi Min-Jung;Kim Jong-Oh;Kim Seok-Ryel;Kim Hyeung-Rak
    • Fisheries and Aquatic Sciences
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    • v.9 no.2
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    • pp.57-63
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    • 2006
  • A bacterial strain capable of hydrolyzing carrageenan was isolated from the coast of Busan in Korea. The isolated strain (HS5322) is aerobic, gram-negative, rod-shaped, and motile. Comparison of the 16S rDNA of the isolate with that of known Pseudomonas sp. showed that sequence similarity was at most 95%, implying that the isolate is a new Pseudomonas species. The organism grew optimally at NaCl concentrations of 2.0 to 2.5%. The optimum temperature and pH for carrageenase production in a 72-h flask culture containing 1% carrageenan was $20^{\circ}C$ and pH 8.5, respectively. Of the synthetic substrates tested, the highest enzyme activity was obtained with p-nitrophenyl ${\beta}$-D-galactopyranoside.

Analysis of antigenic domain of GST fused major surface protein (p30) fragments of Toxoplasma gondii (융합단백질로 발현된 톡소포자충의 주요막단백질(p30) 절편의 항원성)

  • 남호우;임경심
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.135-142
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    • 1996
  • Antigenic domain of jai or surface protein (p30) of Toxoplosmc Sondii was analyzed after polymerase chain reaction (PCR) of its gene fragments. Hydrophilic or hydrophobic moiety of amino acid sequences were expressed as glutathione S-transferase (G57) fusion proteins. Fragments of p30 gene were as follows: 737, total p30 open reading frame (ORF) ; S28, total ORF excluding N-terminal signal sequence and C-terminal hydrophobic sequence; Al9, N-terminal 2/3 parts of A28; A19, N-terminal 2/3 of S28; P9, C-terminal 2/3 part of S28; Z9. middle 1/3 of S28; and 29, C-terminal 1/3 of S28. respectively. Primer of each fragment was synthesized to include clamp sequence of EcoR I restriction site. PCR amplified DNA was inserted info GST (26 kDa) expression vector, PGEX-47-1 to transform into Escheri,hia coei (.JM105 strain). G57 fusion proteins were expressed with IPTG induction as 63. 54, 45, 45, 35, 36. and 35 kDa proteins measured by SDS-PAGE. Each fusion protein was confirmed with G57 detection kit. Western blot analysis with the serum of a toxoplasmosis patient revealed antigenicity in proteins expressed by T37. S28, and Al9 but not those by Pl8. X9, Y10, and Z9. Antigenicity of p30 seems to be located either in N-terminal 115 part in the presence of middle 1/3 part or in the oligopeptides between margins of the first and second 1/3 parts.

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Association of a p53 Codon 72 Gene Polymorphism with Environmental Factors and Risk of Lung Cancer: a Case Control Study in Mizoram and Manipur, a High Incidence Region in North East India

  • Saikia, Bhaskar Jyoti;Das, Mandakini;Sharma, Santanu Kumar;Sekhon, Gaganpreet Singh;Zomawia, Eric;Singh, Yanglem Mohen;Mahanta, Jagadish;Phukan, Rup Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10653-10658
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    • 2015
  • Background: A very high incidence of lung cancer is observed in Mizoram and Manipur, North East India. We conducted a population based case control study to establish associations of p53 codon 72 polymorphisms and interactions with environmental factors for this high incidence. Material and Methods: A total of 272 lung cancer cases and 544 controls matched for age (${\pm}5years$), sex and ethnicity were collected and p53 codon 72 polymorphism genotypes were analyzed using a polymerase chain based restriction fragment length polymorphism assay. We used conditional multiple logistic regression analysis to calculate adjusted odds ratios and 95% confidence intervals after adjusting for confounding factors. Results: p53 Pro/Pro genotype was significantly associated with increased risk of lung cancer in the study population (adjusted OR=2.14, CI=1.35-3.38, p=0.001). Interactions of the p53 Pro/Pro genotype with exposure to wood smoke (adjusted OR=3.60, CI=1.85-6.98, p<0.001) and cooking oil fumes (adjusted OR=3.27, CI=1.55-6.87, p=0.002), betel quid chewing (adjusted OR=3.85, CI=1.96-7.55, p<0.001), tobacco smoking (adjusted OR=4.42, CI=2.27-8.63, p<0.001) and alcohol consumption (adjusted OR=3.31, CI=1.10-10.03, p=0.034) were significant regarding the increased risk of lung cancer in the study population. Conclusions: The present study provided preliminary evidence that a p53 codon 72 polymorphism may effect lung cancer risk in the study population, interacting synergistically with environmental factors.

Association between Social Phobia and Serotonin Transporter Gene Polymorphism : Preliminary Study (사회공포증과 세로토닌 수송체 유전자다형성과의 연관성 : 예비연구)

  • Lee, Jae-Hon;Lim, Se-Won;Oh, Kang-Seob;Lee, Min-Soo
    • Korean Journal of Biological Psychiatry
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    • v.13 no.3
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    • pp.170-177
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    • 2006
  • Objectives : Disturbances of serotonergic system might be related to the possible mechanism of social phobia. This study was to investigate the association of serotonin transporter gene and social phobia. Methods : Sixty nine patients with social phobia(51 male(73.9%), mean age $35.17{\pm}11.89$ years) and seventy four normal controls(54 male(73.0%), mean age $33.46{\pm}9.63$ years) were tested for serotonin transporter gene-linked polymorphic region(5-HTTLPR) polymorphism. Additionally, patients were grouped into 46 generalized(GEN) and 23 nongeneralized(NGEN) subgroups and 5-HTTLPR polymorphism was compared with that of normal controls. The genotypes and allele frequencies of the 5-HTTLPR polymorphism between social phobia and the control group were compared. Genomic DNA was extracted from their blood and 5-HTTLPR polymorphisms were determined by using polymerase chain reaction. Results : Significant association was observed between the S(ss) genotype and social phobia, by functional classification(p=.010). In allele frequency analysis, a significant association was also observed between the short allele and social phobia(p=.030). A significant associations between S genotype and each subgroup were observed(GEN p=.045 ; NGEN p=.033), but there were no differences in allele frequency. And, no differences in genotype and allele distribution between two subgroups were found. Conclusion : The results in our Korean sample suggest that S genotype of 5-HTTLPR may be associated with social phobia and s allele may be an important genetic factor that activates social phobic symptoms. But, further studies including large number of samples are necessary to elucidate these present findings.

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All-trans retinoic acid alters the expression of adipogenic genes during the differentiation of bovine intramuscular and subcutaneous adipocytes

  • Chung, Ki Yong;Kim, Jongkyoo;Johnson, Bradley J.
    • Journal of Animal Science and Technology
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    • v.63 no.6
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    • pp.1397-1410
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    • 2021
  • The present study was designed to determine the influence of all-trans retinoic acid (ATRA) on adipogenesis-related gene regulation in bovine intramuscular (IM) and subcutaneous (SC) adipose cells during differentiation. Bovine IM and SC adipocytes were isolated from three 19-mo-old, crossbred steers. Adipogenic differentiation was induced upon cultured IM and SC preadipocytes with various doses (0, 0.001, 0.01, 0.1, 1 µM) of ATRA. After 96 h of incubation, cells were harvested and used to measure the gene expression of CCAAT/Enhancer binding protein β (C/EBPβ), peroxisome proliferator-activated receptor (PPAR) γ, glucose transporter 4 (GLUT4), stearoyl CoA desaturase (SCD), and Smad transcription factor 3 (Smad3) relative to the quantity of ribosomal protein subunit 9 (RPS 9). Retinoic acid receptor (RAR) antagonist also tested to identify the effect of ATRA on PPARγ -RAR related gene expression in IM cells. The addition of ATRA to bovine IM decreased (p < 0.05) expression of PPARγ. The expression of PPARγ was also tended to be downregulated (p < 0.1) in high levels (10 µM) of ATRA treatment in SC cells. The treatment of RAR antagonist increased the expression of PPARγ in IM cells. Expression of C/EBPβ decreased (p < 0.05) in SC, but no change was observed in IM (p > 0.05). Increasing levels of ATRA may block adipogenic differentiation via transcriptional regulation of PPARγ. The efficacy of ATRA treatment in adipose cells may vary depending on the location.

Expression of in vitro-tested ribozyme against cucumber mosaic virus RNA in tobacco plant (시험관내에서 합성한 오이모자이크 바이러스 RNA단편을 성공적으로 절단한 ribozyme의 식물체내의 발현)

  • Park, Sang-Gyu
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.355-360
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    • 1996
  • In vitro-tested ribozyme against synthesized cucumber mosaic virus (CMV) RNA (Agric. Chem. & Biotech. 37:56-63(1994)) was expressed in tobacco plant to develop virus resistant plants. The ribozyme sequence was linked to cauliflower mosaic virus 35S promoter and nopaline synthase(nos) terminator and this chimeric 35S-ribozyme-nos gene was sequenced. The sequenced chimeric gene was transferred to Agrobacterium tumefaciens LBA4404 using tri-parental mating system. The E. coli HB101 containing chimeric gene was incubated with E. coli HB101(pRK2073) as a helper and Agrobacterium tumefaciens LBA4404. Then Agrobacterium cells containing the ribozyme construct was cocultivated with tobacco leaf pieces. Ten different plants were regenerated from kanamycin containing MS medium. The presence of the ribozyme construct in the transgenic tobacco plants was confirmed by polymerase chain reaction (PCR). Seven different transgenic plants in ten different kanamycin resistant plants showed the expected size (570 base pairs) of 35S-ribozyme-nos gene fragment. Total RNAs were isolated from four different transgenic plants and separated on a 1% agarose gel containing formamide. Northern hybridization with 35S-ribozyme-nos gene fragment as a probe indicated that ribozyme transcripts may be degraded tv nuclease. Therefore, nuclease-resistant ribozymes are needed for the development of virus-resistant transgenic plants using ribozymes.

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Co-expression and Sequence Determination of Estrogen Receptor Variant Messenger RNAs in Swine Uterus

  • Ying, C.;Chan, M.-A.;Cheng, W.T.K.;Hong, W.-F.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1716-1721
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    • 2003
  • Steroid hormones and their receptors play an important role in reproductive process. Estrogen is intimately involved with pregnancy and its function is mediated through the estrogen receptor which has been chosen as a candidate gene to study litter size in pigs. In this study, we report that two estrogen receptor variants, designated pER-1 and pER-2 were co-expressed in the uteri of normal cycling Lan-Yu pig (Sus vittatus; a small-ear miniature in Taiwan) with the pER-1 expression level appeared to be several times higher than that of pER-2. These receptor variants were isolated using reverse transcription-PCR from the pig uteri and their sequences were determined. The pER-1 and pER-2 sequences, which are homologous to those found in other mammalian estrogen receptors, encode putative proteins consisting of 574 and 486 amino acids, respectively. A deletion in exon I was identified in both sequences, with deletion lengths of 63 bp in pER-1 and 327 bp in pER-2. The deletion in pER-1 is internal to that in pER-2 and both deletions resulted in a truncation of the B domain, which confers the transactivating activity of estrogen receptor protein. This result describes the existence of estrogen receptor variants with a deletion in exon I and implies the possibility that physiological functioning of an estrogen receptor may not require the presence of an intact B domain.

Clinical Characteristics and Prognosis of Differentiated Thyroid Carcinoma with Small Foci of Anaplastic Transformation

  • Ahn, Hwa Young;Jung, Kyeong Choen;Park, Do Joon;Park, Young Joo;Cho, Bo Youn
    • International journal of thyroidology
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    • v.10 no.2
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    • pp.96-101
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    • 2017
  • Background and Objectives: Anaplastic thyroid carcinoma (ATC) is commonly related with concurrent differentiated thyroid carcinoma (DTC). We aimed to examine the clinicopathologic characteristics, prognosis and gene expression of DTC with anaplastic foci. Materials and Methods: Eighteen patients with DTC with anaplastic foci were enrolled in this study. To compare the clinicopathologic characteristics and prognosis of anaplastic foci subjects with conventional ATC or DTC, we additionally included 12 ATC and 1030 DTC patients who diagnosed during same period. Immunohistochemistry was performed to check the gene expression in anaplastic foci and DTC component. Results: In anaplastic foci group, tumor size was larger ($2.5{\pm}1.3$ vs. $1.2{\pm}0.9cm$, p=0.001), distant metastasis was more frequent (11.1 vs. 0%, p=0.000) and 1-year survival rate was low (88.9 vs. 100%, p=0.000) than DTC group. In contrast, compared with ATC group, anaplastic foci group showed younger age at diagnosis ($50{\pm}16$ vs. $63{\pm}18years$, p=0.039), smaller tumor size ($2.5{\pm}1.3$ vs. $3.8{\pm}1.4cm$, p=0.027), less distant metastasis (11.1 vs. 41.7%, p=0.084) and longer 1-year survival rate (88.9 vs. 25.0%, p=0.001). Expression of p53 protein was observed in 100% of anaplastic foci, ATC and 12.5% of papillary thyroid carcinoma component. Conclusion: DTC with foci of anaplastic transformation has a worse prognosis than DTC, but a better prognosis than ATC. Our results support that DTC with anaplastic foci was intermediate state from DTC to ATC.

The Effects of Triallelic Serotonin Transporter Gene Polymorphism and Stressful Life Event on Depression in Patients with Alcohol Dependence (알코올 의존 환자에서 삼대립 세로토닌 수송체 유전자 다형성과 생활사건 스트레스가 우울증에 미치는 영향)

  • Jang, Hyun-Chung;Lee, Sang-Ick;Kim, Sie-Kyeong;Shin, Chul-Jin;Son, Jung-Woo;Ju, Ga-Won;Park, Jae-Young;Jee, Kyung-Hwan;Lee, Sang-Gu
    • Korean Journal of Biological Psychiatry
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    • v.19 no.2
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    • pp.106-113
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    • 2012
  • Objectives : The purpose of this study is to investigate the relationship between the triallelic serotonin transporter gene and stressful life events to determine their effect on depression with alcohol dependence. Methods : Ninety-five hospitalized patients with alcohol dependence (73 male, 22 female) were enrolled in this study. Thirty-two (33.7%) of the total patients were diagnosed with major depressive disorder and dysthymic disorder by Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders-IV. The characteristics of stress were evaluated using the stressful life events scale, and depressive symptoms were assessed using the depression scale (Beck Depression Inventory, BDI). Alcoholism with depression (n = 32) and alcoholism without depression (n = 63) were genotyped for the triallelic serotonin transporter gene ($L_A$ : higher expressing allele, $L_G$/S : lower expressing allele). Results : There was no significant difference in the allele frequency between the depression group and the non-depression group (${\chi}^2$ = 0.345, p = 0.619). $L_G$/S alleles had more comorbid depression in the higher score of stressful life events scale [Mental-Haenszel (MH)-${\chi}^2$ = 4.477, p = 0.034]. But there was no significant difference in the comorbidity according to the scores from the stressful life event scale in the $L_A$ alleles (MH-${\chi}^2$ = 0.741, p = 0.399). In the results, alcohol-dependent individuals with $L_G$/S alleles had more comorbid depression than those with $L_A$ alleles when they had experienced severe stressful life events (MH-odds ratio = 2.699, p = 0.028). Conclusions : These results suggest that there is no direct relationship between triallelic serotonin transporter gene and depression in the alcohol dependent patients. But alcohol dependent individuals with the lower expressing alleles of the serotonin transporter gene were more susceptible to depression than those with the higher expressing alleles in response to stressful life events.

Carrier frequency of SLC26A4 mutations causing inherited deafness in the Korean population

  • Kim, Hyogyeong;Lim, Hwan-Sub;Ryu, Jae-Song;Kim, Hyun-Chul;Lee, Sanghoo;Kim, Yun-Tae;Kim, Young-Jin;Lee, Kyoung-Ryul;Park, Hong-Joon;Han, Sung-Hee
    • Journal of Genetic Medicine
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    • v.11 no.2
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    • pp.63-68
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    • 2014
  • Purpose: The mutation of the SLC26A4 gene is the second most common cause of congenital hearing loss after GJB2 mutations. It has been identified as a major cause of autosomal recessive nonsyndromic hearing loss associated with enlarged vestibular aqueduct and Pendred syndrome. Although most studies of SLC26A4 mutations have dealt with hearing-impaired patients, there are a few reports on the frequency of these mutations in the general population. The purpose of this study was to evaluate the prevalence of SLC26A4 mutations that cause inherited deafness in the general Korean population. Materials and Methods: We obtained blood samples from 144 Korean individuals with normal hearing. The samples were subjected to polymerase chain reaction to amplify the entire coding region of the SLC26A4 gene, followed by direct DNA sequencing. Results: Sequencing analysis of this gene identified 5 different variants (c.147C>G, c.225G>C, c.1723A>G, c.2168A>G, and c.2283A>G). The pathogenic mutation c.2168A>G (p.H723R) was identified in 1.39% (2/144) of the subjects with normal hearing. Conclusion: These data provide information about carrier frequency for SLC26A4 mutation-associated hearing loss and have important implications for genetic diagnostic testing for inherited deafness in the Korean population.