• Title/Summary/Keyword: E6 and E7 genes

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HEMAGGLUTINATION AND COLONY HYBRIDIZATION FOR THE IDENTIFICATION OF ENTEROTOXIGENIC Escherichia coli ISOLATED FROM HEALTHY PIG

  • Choi, S.H.;Oh, M.J.;Sung, C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.9 no.6
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    • pp.671-677
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    • 1996
  • Erythrocytes from three different animal species were used to determine mannose-sensitive hemagglutination (MSHA) and mannose-resistant hemagglutination (MRHA) of 755 isolates obtained from rectal swabs of healthy pig. In addition, colony hybridization using digoxigenin-dUTP labeled polynucleotide probes was performed for the detection of heat-stable and heat-labile enterotoxin genes carried by MRHA positive isolates. Of 755 strains, 9, 4 and 28 strains gave a positive MRHA with bovine, equine and pig erythrocytes, respectively. Of these isolates, 28 (3.7%) were characterized for positive MRHA by at least one blood. Seven isolates gave a positive MRHA with two kinds of blood. Three gave a positive MRHA with three kinds of blood. Twenty-eight strains, while positive in MRHA, yielded negative signals in the colony hybridization assay for the detection of heat-stable (STaI and STaII) and heat-labile (LT) enterotoxin genes in E. coli.

Replicated Association Study between Tuberculosis and CLCN6, DOK7, HLA-DRA in Korean

  • Kim, Sung-Soo;Park, Min;Park, Sangjung
    • Biomedical Science Letters
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    • v.26 no.3
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    • pp.238-243
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    • 2020
  • Tuberculosis is a global public health problem and manifests itself as a difference in the genetic susceptibility of the host, along with the properties of Mycobacterium tuberculosis (MTB). The single nucleotide polymorphisms (SNPs) and candidate genes proposed in the Genome-wide association study (GWAS) on tuberculosis in a recently published Chinese population were reported. In this study, we investigated whether the genetic polymorphism of candidate genes related to tuberculosis is reproduced when targeting Koreans. The CLCN6 (rs12404124, rs198391, rs535107), DOK7 (rs1203104, rs1203103) and HLA-DRA (rs1051336) gene polymorphisms showed statistically significant results. In addition, it was also found whether it acts as an expression quantitative trait loci (eQTL) that can influence gene expression. This study confirmed that the genetic polymorphism of the three genes (CLCN6, DOK7, HLA-DRA) affects the development of tuberculosis and will help to understand the genetic specificity of tuberculosis and the interaction between pathogens and hosts.

Identification of Genes Differentially Expressed in the MCF-7 Cells Treated with Mitogenic Estrogens

  • Cheon, Myeong-Sook;Yoon, Tae-Sook;Lee, Do-Yeon;Choi, Go-Ya;Lee, A-Yeong;Choo, Byung-Kil;Kim, Ho-Kyoung
    • Journal of Applied Biological Chemistry
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    • v.51 no.1
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    • pp.1-6
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    • 2008
  • Estrogens, a group of steroid compounds functioning as the primary female sex hormone, play an important role in the development and progression of breast cancer. In this study, using a novel annealing control primer-based GeneFishing PCR technology, five differentially expressed genes (DEGs), expressed using 10nM mitogenic estrogens, $17{\beta}$-estradiol (E2) and $16{\alpha}$-hydroxyestrone ($16{\alpha}$-OHE1), were selected from the estrogen receptor (ER)-positive MCF-7 human breast cancer cells. The DEGs, MRPL42, TUBA1B, SSBP1, KNCT2, and RUVBL1, were identified by comparison with the known genes via direct sequencing and sequence homology search in BLAST. Quantitative real-time PCR data showed that two DEGs, tubulin ${\alpha}1b$ and kinetochore associated 2, were greater than 2-fold upregulated by E2 or $16{\alpha}$-OHE1. Both genes could be new biomarkers for the treatment and prognosis of cancers, and further study may provide insights into the molecular mechanisms underlying development and progression of breast cancer.

Characterization of G9 genotype porcine rotavirus isolated in Korea (국내 양돈장에서 분리한 G9형 돼지로타바이러스의 특성)

  • Wang, Jun-Hui;Lee, Seung-Chul;Kang, Shien-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.2
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    • pp.83-90
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    • 2012
  • Porcine rotaviruses are the most common causes of viral gastroenteritis in piglets around the world. The major G genotypes of porcine rotaviruses causing diarrhea were G4, G5 and G11 genotypes. Recently, G9 genotype rotaviruses were problemed at swine farms and frequently recognized from diarrheic piglets. In this study, a porcine rotavirus (PoRV-1) was isolated from piglet showing diarrhea using MA104 cells and confirmed as rotavirus by electron microscopy, genomic RNA electropherotyping and indirect immunofluorescence antibody tests. The nucleotide sequence of the VP7 gene of PoRV-1 was determined and compared with those of other genotype rotavirus strains from other parts of the world. Also, the nucleotide sequences of VP4, VP6 and NSP4 genes of PoRV-1 were determined and compared with those of other rotavirus strains from other countries. The results showed that the PoRV-1 isolate belonged to the G9 genotype and the P, I and E genotypes of PoRV-1 were P[23], I5 and E1, respectively. The Korean G9 PoRV-1 isolate and its nucleotide sequence data would be usefully used for the development of porcine rotavirus vaccines in near future.

Genetics of Alzheimer's Disease

  • Kim, Jong Hun
    • Dementia and Neurocognitive Disorders
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    • v.17 no.4
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    • pp.131-136
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    • 2018
  • Alzheimer's disease (AD) related genes have been elucidated by advanced genetic techniques. Familial autosomal dominant AD genes founded by linkage analyses are APP, PSEN1, PSEN2, ABCA7, and SORL1. Genome-wide association studies have found risk genes such as ABCA7, BIN1, CASS4, CD33, CD2AP, CELF1, CLU, CR1, DSG2, EPHA1, FERMT2, HLA-DRB5-HLA-DRB1, INPP5D, MEF2C, MS4A6A/MS4A4E, NME8, PICALM, PTK2B, SLC24A4, SORL1, and ZCWPW1. ABCA7, SORL1, TREM2, and APOE are proved to have high odds ratio (>2) in risk of AD using next generation sequencing studies. Thanks to the promising genetic techniques such as CRISPR-CAS9 and single-cell RNA sequencing opened a new era in genetics. CRISPR-CAS9 can directly link genetic knowledge to future treatment. Single-cell RNA sequencing are providing useful information on cell biology and pathogenesis of diverse diseases.

Inhibition of adhesion and biofilm formation in Escherichia coli O157:H7 by diosmin (다이오스민(diosmin)에 의한 병원성 대장균 세포부착 및 생물막 형성 억제)

  • Kim, Hyun Jung;Kim, Seung Min
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.414-419
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    • 2018
  • Escherichia coli O157:H7 is one of the most common foodborne pathogens responsible for outbreaks of hemorrhagic colitis, which can lead to the life-threatening hemolytic-uremic syndrome. In this study, we identified phytochemicals that specifically inhibit the expression of LEE operon in E. coli O157:H7. Among phytochemicals, diosmin decreased the adherence of E. coli O157:H7 towards Caco-2 cells in vitro (p<0.01) and its biofilm formation activity (p<0.05). Quantitative RT-PCR analysis revealed that the transcripts of Ler-regulated genes and genes related to curli production were significantly reduced in the presence of diosmin. However, diosmin does not affect bacterial viability, indicating that the resistance rate to diosmin was remarkably low. Overall, these results provide significant insights into the development of a novel anti-infective agent that is different from conventional antibiotics.

Detection and Typing of Human Papillomavirus in Cutaneous Common Warts by Multiplex Polymerase Chain Reaction (Multiplex PCR 기법을 이용한 보통사마귀 내 인유두종바이러스 검출 및 분류)

  • Choi, Soon-Yong;Lim, Jong-Ho;Kim, Eun-Jung;Kim, Hei-Sung;Kim, Beom-Joon;Kang, Hoon;Park, Young-Min
    • Journal of Life Science
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    • v.21 no.7
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    • pp.947-952
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    • 2011
  • A number of epidemiological studies have identified human papillomavirus (HPV) types 1, 2, 3, 4, 7, 10, 27, 57, and 65 in cutaneous common warts. However, identification of the HPV subtype by conventional polymerase chain reaction (PCR) is time consuming with its multi-step laboratory process. In this study, we aim to develop a specific one-step multiplex polymerase chain reaction method which capably identifies six different HPV genotypes related to common warts. By HPV DNA sequence analysis, 6 pairs of specific primers were designed from the intergenic regions of genes L1 to E6, and from genes E2 to L2. DNA sequence analysis with the L1 gene sequence of the sample was performed to measure the specificity of multiplex PCR. HPV-1, -2, -3, -4, -27, and -57 were identified without cross amplification in 109 out of 129 samples. The sensitivity and specificity of our set of primers in detecting HPV were 85% and 99.5%, respectively. For the 20 samples where HPV type was not identifiable by our batch of primer sets, multiplex PCR with an additional set of HPV primers was done, where 7 were found positive for HPV-7 or -65. Our results demonstrate that the newly designed multiplex PCR can rapidly detect the specific HPV subtype involved in common warts with high accuracy.

A New Deoxyhexose Biosynthetic Gene Cluster in Streptomyces griseus ATCC10137: Heterologous Expression of dTDP-D-Glucose 4,6-Dehydratase Gene

  • Kim, Sang Suk;Bang, Jung-Hee;Hyun, Chang-Gu;Kim, Joo-Woo;Han, Jae-Jin;Suh, Joo-Won
    • Journal of Applied Biological Chemistry
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    • v.43 no.3
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    • pp.136-140
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    • 2000
  • A novel 6-deoxyhexose biosynthetic gene cluster different from the one for the biosynthesis of streptomycin was isolated from Streptomyces griseus using specifically designed PCR primers to compare the sequence of known dTDP-glucose synthase genes. We cloned a 5.8-kb DNA from Streptomyces griseus ATCC10137, which contained the 4-ketoreductase homologue (grsB), dTDP-glucose synthase (grsD), and dTDP-glucose 4, 6-dehydratase (grsE) genes. Escherichia coli cultures containing plasmid of the PCR product which encoded the grsE region under the controUed T7 promoter were able to catalyze the formation of dTDP-4-keto-6-deoxy-D-glucose from TDP-glucose. The enzyme showed high substrate specificity, being specific to only dTDP-glucose that is known to be incorporated into secondary metabolites such as antibiotics.

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Mutation Detection of E6 and LCR Genes from HPV 16 Associated with Carcinogenesis

  • Mosmann, Jessica P.;Monetti, Marina S.;Frutos, Maria C.;Kiguen, Ana X.;Venezuela, Raul F.;Cuffini, Cecilia G.
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.1151-1157
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    • 2015
  • Human papillomavirus (HPV) is responsible for one of the most frequent sexually transmitted infections. The first phylogenetic analysis was based on a LCR region fragment. Nowadays, 4 variants are known: African (Af-1, Af-2), Asian-American (AA) and European (E). However the existence of sub-lineages of the European variant havs been proposed, specific mutations in the E6 and LCR sequences being possibly related to persistent viral infections. The aim of this study was a phylogenetic study of HPV16 sequences of endocervical samples from C${\acute{o}}$rdoba, in order to detect the circulating lineages and analyze the presence of mutations that could be correlated with malignant disease. The phylogenetic analysis determined that 86% of the samples belonged to the E variant, 7% to AF-1 and the remaining 7% to AF-2. The most frequent mutation in LCR sequences was G7521A, in 80% of the analyzed samples; it affects the binding site of a transcription factor that could contribute to carcinogenesis. In the E6 sequences, the most common mutation was T350G (L83V), detected in 67% of the samples, associated with increased risk of persistent infection. The high detection rate of the European lineage correlated with patterns of human migration. This study emphasizes the importance of recognizing circulating lineages, as well as the detection of mutations associated with high-grade neoplastic lesions that could be correlated to the development of carcinogenic lesions.

Conditions for Stable light Production of Recombinant Escherichia coli Containing Lux Operon and Sensitivity to Toxic Chemicals (Lux operon을 함유한 유전자 재조합 Escherichia coli의 발광 안정화 조건 및 독성물질에 대한 민감성)

  • 배희경;이상민;정윤철;송방호;신평균
    • KSBB Journal
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    • v.17 no.6
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    • pp.571-576
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    • 2002
  • Recombinant E. coli DH5 ${\alpha}$/pSB311 was made by cloning the genes encoding bacterial luciferase and aldehyde substrate proteins from Photohabdus luminescense, to complement defects of Lumistox, which is normally used in bioassays to monitor toxic substances in water environmental systems. The conditions for stable light production by the recombinant strains were investigated with respect to cell growth stage, cell number, and buffer conditions. The optimum growth stage was a middle-exponential stage with an OD$_{660nm}$ value of 0.6-0.7. ADout 10$^{6}$-10$^{7}$ cells per test tube was optimum for stable light emission. The effect of buffer was not significant if an optimum viable cell number was maintained. The bioluminescence of the recombinant E. coli harboring the lux operon of Photohabdus luminescense was not affected by temperature, while the bioluminescence of Lumistox was temperature sensitive. The recombinant E. coli was more sensitive to heavy metals (Cd, Cu, Hg, Zn) than Lumistox, because it does not require high concentrations of NaCl in the buffer.